TWI911619B - Fermentation method for enhancing volatility material of vanilla seed pod with bacillus velezensis and product produced thereby - Google Patents

Abstract

A vanilla seed pod fermentation method includes: prefabricating at least one fungi bacillus velezensis; blanching a vanilla seed pod with a high-temperature blanching procedure to obtain a blanched vanilla seed pod; adding the fungi bacillus velezensis to the blanched vanilla seed pod for fungi-fermenting the blanched vanilla seed pod; dehydrating, fungi-fermenting and sterilizing the blanched Vanilla seed pod to obtain a dehydrated, fermented Vanilla seed pod to obtain a volatility-fermenting-enhanced Vanilla seed pod; and drying the volatility-fermentation-enhanced Vanilla seed pod in a high-temperature roasting procedure to obtain a dried, volatility-fermentation-enhanced Vanilla seed pod.

Description

A method for enhancing the fermentation of volatile compounds in orchid pods using bacillus velezensis and the resulting products.

This invention relates to a method for enhancing fermentation of the volatile material of vanilla seed pod using bacillus velezensis , and to products thereof; particularly, to a method for enhancing fermentation of the volatile material of vanilla seed pod using bacillus velezensis at a suitable temperature (e.g., room temperature), and to products thereof.

Regarding commonly used low-temperature fermentation methods for sago palm pods, for example, the Republic of China Patent Publication No. TW-202327468, "Fermentation Method and Materials Thereof," discloses a low-temperature fermentation method and materials for sago palm pods. This low-temperature fermentation method and materials for sago palm pods can optionally employ a low-temperature fermentation system.

Continuing from the above, the low-temperature fermentation method for pods of *Calvatia* as disclosed in Patent Publication No. TW-202327468 comprises: subjecting a pod of *Calvatia* to a high-temperature blanching process to obtain a blanched pod; subjecting the blanched pod of *Calvatia* directly to a low-temperature dehydration process, a low-temperature fermentation process, and a sterilization process to obtain a dehydrated and fermented pod; and subjecting the dehydrated and fermented pod of *Calvatia* to a high-temperature drying process to obtain a dried and fermented pod.

Continuing from the above, the low-temperature fermentation method for pods of *Cymbidium goeringii* as disclosed in Patent Publication No. TW-202327468 comprises: subjecting the sterilized pods of *Cymbidium goeringii* to low-temperature fermentation catalysis at a predetermined temperature using a solar lamp catalytic unit, wherein the predetermined temperature includes a range of 15°C to 25°C or a related temperature range, and the low-temperature dehydration, low-temperature fermentation, or sterilization is selectively performed in a refrigerated drying oven unit.

Continuing from the above, the low-temperature fermentation method for pods of senna, as disclosed in Patent Publication No. TW-202327468, comprises: drying the dehydrated and fermented pods of senna at a predetermined temperature, wherein the predetermined temperature is between 30°C and 100°C or a related temperature range.

Another commonly used method for rapid fermentation of pods is exemplified by the Republic of China Patent No. TW-I756953, "Rapid Fermentation Method of Piper Pods," which discloses a rapid fermentation method for pods. This rapid fermentation method includes step one: treating a harvested pod with a blanching process at 60°C to 65°C for 1 to 10 minutes to obtain a blanched pod.

Continuing from the above, the rapid fermentation method for pods of *Callicarpa stolonifera* as described in the aforementioned patent announcement TW-I7569532 includes step two: fermenting the blanched pods at a temperature of 40°C and a humidity of 40% to 60% for approximately two days to obtain fermented pods.

Continuing from the above, the rapid fermentation method for pods of *Callicarpa japonica* as described in the aforementioned patent announcement TW-I7569532 includes step three: placing the fermented pods in a drying oven and performing a drying process for approximately 4 to 6 weeks at a temperature of 40°C and a relative humidity of 40% to 50% to rapidly dry the fermented pods and obtain dried pods; wherein a desiccant is provided inside the drying oven.

Continuing on, the rapid fermentation method for senna pods in the aforementioned patent announcement No. TW-I7569532 includes step four: the dried senna pods are placed in the drying box and subjected to a first-maturation process for about 2 to 4 months at a temperature of 40°C and a relative humidity of 40 to 50% to obtain first-matured senna pods. Continuing from the above, the rapid fermentation method for pods of *Callicarpa stolonifera* as described in the aforementioned patent announcement TW-I7569532 includes step four: The dried pods are further placed in the drying chamber and subjected to a first-maturation process for approximately 2 to 4 months at a temperature of 40°C and a relative humidity of 40% to 50% to obtain first-matured pods.

However, the rapid fermentation method for senna pods described in the aforementioned patent announcement TW-I7569532 requires fermentation, drying, and maturation to be carried out under relatively high temperature conditions (e.g., 40°C). Therefore, it suffers from the disadvantages of operating at relatively high temperatures and consuming relatively high energy. Furthermore, it requires placement in various different devices (e.g., drying ovens) for fermentation, drying, and maturation, making it susceptible to contamination (e.g., bacterial infection) during frequent equipment changes.

Another commonly used method is vanilla seed fermented milk, its formula, and its preparation method. For example, Chinese Patent Publication No. CN-102626142, "Vanilla Seed Fermented Milk, Its Formula, Preparation Method, and Semi-Quantitative Analysis Method," discloses a vanilla seed fermented milk, its raw material formula, and its preparation method.

Continuing from the above, the method for preparing vanilla seed fermented milk disclosed in patent CN-102626142 includes step 1: mixing the fermented milk with vanilla seed jam with a viscosity not exceeding 10 cm/s [20°C, 60 seconds] and filling; during the filling process, a plunger pump of model DACC48/100-23 from PCM (France) is used when adding the jam by metering.

Continuing on, the method for preparing vanilla seed fermented milk disclosed in the aforementioned patent disclosure CN-102626142 further includes, in step 1: the fermented milk is obtained by conventional methods for preparing fermented milk, and the raw material formula of the vanilla seed fermented milk includes raw milk, milk protein powder, milk fat products, sweeteners, thickeners, fermenting bacteria, and vanilla seed jam with a viscosity not exceeding 10 cm/s [20°C, 60 seconds].

Continuing from the above, the method for preparing vanilla seed fermented milk according to the aforementioned patent disclosure CN-102626142 includes step 2: using a semi-quantitative analysis method to determine the amount of jam added in the vanilla seed fermented milk, and selecting vanilla seed fermented milk that meets the target amount.

However, the aforementioned patent disclosure CN-102626142 merely discloses the vanilla seed fermented milk and its preparation method, as well as the application technology of adding vanilla seed jam to it. It clearly does not further disclose how to use appropriate technical means to carry out low-temperature fermentation of vanilla pods, its processing methods, or related technical concepts.

Another common method involves preparing tobacco flakes containing black stalk tobacco and its preparation process. For example, Chinese Patent Publication No. CN-103445286, "A Method for Preparing Tobacco Flakes Containing Black Stalk Tobacco," discloses a method for preparing tobacco flakes containing black stalk tobacco, which is applied in the tobacco manufacturing process.

Continuing from the above, the method for preparing the black stalk tobacco flakes disclosed in Patent Publication No. CN-103445286 includes step 1: removing some of the oil from the black stalk tobacco, then drying the partially oil-removed black stalk tobacco at 72°C to 78°C to obtain dried black stalk tobacco, and further removing some of the oil from the black stalk tobacco in ether.

Continuing from the above, the method for preparing black stalk tobacco flakes disclosed in Patent Publication No. CN-103445286 includes step 2: first, vanillin and ethyl vanillin are prepared into solutions with a weight percentage of 75% and 25% respectively in propylene glycol, and mixed to obtain a mixed solution; then, the mixed solution with a weight of 1% to 3% of the wet weight of black stalk tobacco is uniformly sprayed onto the surface of dried stalk tobacco, and then dried at 28°C to 32°C; finally, the dried stalk tobacco is pulverized and sieved to obtain processed black stalk tobacco powder.

Continuing from the above, the preparation method of the black stalk tobacco flakes disclosed in Patent Publication No. CN-103445286 includes step 3: adding the treated black stalk powder to the coating liquid at 0.5% to 3% of the total weight of the coating liquid to prepare black stalk tobacco flakes, wherein the coating liquid is selected from a concentrated mixture of tobacco dust, tobacco stems, and tobacco leaf extracts after flue-cured tobacco fermentation.

However, the aforementioned patent disclosure CN-103445286 merely discloses the preparation method of the tobacco flakes containing black stalk orchid and the technology of tobacco preparation methods; it clearly does not further disclose how to use appropriate technical means to carry out low-temperature fermentation of stalk orchid material, its processing methods, or other related technical concepts.

Another commonly used method for fermenting vanilla pods is illustrated in Chinese Patent Publication No. CN-104630288, "A Fermentation Method for Vanilla Pods," which discloses a fermentation method applicable to the fermentation of fresh vanilla pods.

Continuing from the above, the fermentation method for vanilla pods disclosed in Patent No. CN-104630288 includes step 1: washing fresh vanilla pods to obtain washed vanilla pods, and then sequentially performing blanching, sweating, and drying operations to obtain dried vanilla pods. The blanching process involves placing the washed vanilla pods in a water bath at 70±3℃ for 3 to 5 minutes to obtain blanched vanilla pods, and the sweating process involves placing the blanched vanilla pods in a 50℃ oven for 5 days.

Continuing from the above, the fermentation method for vanilla pods disclosed in patent CN-104630288 includes step 2: disinfecting the surface of dried vanilla pods, then uniformly spraying a bacterial suspension with preservation number CGMCC No. 8629 onto the surface, and then aging it in a vacuum-sealed manner to obtain aged vanilla pods.

However, the aforementioned patent disclosure CN-104630288 merely discloses the fermentation method of the vanilla pod, the aged vanilla pod product, and the technology of using a microbial suspension. It clearly does not further disclose how to conduct low-temperature fermentation of vanilla pods using appropriate technical means, its processing methods, or related technical concepts.

Another commonly used method for processing vanilla beans is illustrated by U.S. Patent Application No. US-2011/0081448, "PROCESS FOR TREATING VANILLA BEANS," which discloses a process for processing vanilla beans, specifically for the blanching, crushing, and fermentation of vanilla beans.

Continuing from the above, the vanilla bean processing method disclosed in the aforementioned patent disclosure US-2011/0081448 includes: Step 1, blanching a vanilla bean to obtain a blanched vanilla bean; Step 2, selectively cooling the blanched vanilla bean to obtain a cooled blanched vanilla bean; Step 3, comminuting the cooled blanched vanilla bean to obtain vanilla bean fragments.

Continuing on, the vanilla bean processing method disclosed in the aforementioned patent disclosure US-2011/0081448 includes: Step 4, incubating vanilla bean fragments to obtain fermented vanilla bean fragments, wherein the fermentation is carried out under anaerobic conditions to enhance the hydrolysis of enzymatic glucosides.

Continuing on, the vanilla bean processing method disclosed in the aforementioned patent disclosure US-2011/0081448 includes: Step 5, drying the fermented and cultured vanilla bean fragments to obtain a dried fermented and cultured vanilla bean fragment.

However, the aforementioned patent disclosure No. US-2011/0081448 merely discloses the processing procedure for sago palm beans and the technology of using fermentation cultivation; it clearly does not further disclose how to conduct low-temperature fermentation of sago palm pods using appropriate technical means, its processing methods, or related technical concepts.

Clearly, there is a potential need to further develop methods for low-temperature fermentation of vanilla pods using appropriate technologies, as outlined in the aforementioned Republic of China patents TW-202327468 and TW-I756953, Chinese patents CN-102626142 (method for preparing vanilla seed fermented milk), CN-103445286 (method for preparing materials containing black stalk tobacco flakes), CN-104630288 (fermentation method for vanilla pods), and US patent US-2011/0081448 (method for processing vanilla pods). This would provide advantages such as reduced manufacturing costs, lower energy consumption, simplified overall processes, and standardized industrial production processes.

In short, the technologies disclosed in the aforementioned Republic of China Patent Publication No. TW-202327468, Announcement No. TW-I756953, Chinese Patent Publication Nos. CN-102626142, CN-103445286, CN-104630288, and US Patent Publication No. US-2011/0081448 are merely for reference and illustration of the current state of technological development and are not intended to limit the scope of this invention.

In view of this, in order to meet the above-mentioned technical problems and needs, the present invention provides a method and product for promoting the fermentation of volatile substances in pods of * Bacillus velezensis * using *Bacillus velezensis*. The method involves subjecting a pod of *Bacillus velezensis* to a high-temperature blanching process to obtain a blanched pod, and then directly subjecting the blanched pod to a dehydration process, a microbial fermentation process, and a sterilization process to obtain a fermented pod of *Bacillus velezensis*. The pods of the orchid are subjected to a high-temperature drying process to obtain dried, fermented orchid pods. Therefore, compared with the conventional orchid pod fermentation method, it can achieve the goals of providing fermentation-enhancing volatile substances, reducing manufacturing costs, reducing process energy consumption, simplifying the overall process, and standardizing industrial production processes.

The main objective of this invention is to provide a method for enhancing the fermentation of volatile substances in pods of * Bacillus velezensis* using bacillus velezensis , and the resulting products. The method involves preparing at least one strain of bacteria, subjecting a pod of pods to a high-temperature blanching process to obtain blanched pods, and then incorporating the bacillus strain into the blanched pods to enhance fermentation. The bacillus velezensis is then directly subjected to… The process involves a dehydration process, a microbial fermentation process, and a sterilization process to obtain an enhanced fermented sago palm pod. The enhanced fermented sago palm pod is then subjected to a high-temperature drying process to obtain a dried enhanced fermented sago palm pod. This process aims to provide enhanced fermentation volatiles, reduce manufacturing costs, reduce process energy consumption, simplify the overall process, and achieve standardized industrial production processes.

To achieve the above objectives, a preferred embodiment of the present invention provides a method for promoting the enhanced fermentation of volatile substances in sago palm pods using *Bacillus velezensis* , comprising: preparing at least one strain selected from an enhanced fermentation strain; subjecting a sago palm pod to a high-temperature purifying process to obtain a purified sago palm pod; and combining the strain with the purified sago palm pod to enhance the fermentation of the purified sago palm pod using the strain. The pod is subjected to a dehydration process, a microbial fermentation process, and a sterilization process to obtain an enhanced fermented pod containing at least one volatile substance and at least one vanillin; and the enhanced fermented pod is subjected to a high-temperature drying process to obtain a dried enhanced fermented pod.

In a preferred embodiment of the present invention, the strain may be selected from bacillus velezensis or bacillus velezensis ZN-S10.

The preferred embodiment of this invention has a sequence, which can be selected from Seq.1.

In a preferred embodiment of the present invention, the volatile substance forms an aroma compound (e.g., alcohols, ketones, phenols, aldehydes, esters, terpenes, hydrocarbons, or carboxylic acids).

In a preferred embodiment of the present invention, the cyanided vanilla pod is fermented using microorganisms to adjust and form the volatile substances and their aroma.

In a preferred embodiment of the invention, the high-temperature sterilization operation is performed in a high-temperature steam unit or an insulated container unit.

In a preferred embodiment of the invention, the dehydration, microbial fermentation, or sterilization process is performed in a refrigerated drying unit.

In a preferred embodiment of the invention, the dehydration, fermentation, and sterilization processes are selectively performed within a single refrigerated drying unit.

In a preferred embodiment of the invention, the high-temperature drying operation is performed in an oven unit.

To achieve the above objectives, a preferred embodiment of the present invention, a pod-based product of pods that uses bacillus velezensis to promote enhanced fermentation and the formation of volatile substances, comprises: at least one strain selected from a fermentation-enhancing strain , bacillus velezensis. ; and at least one pod of sambac, wherein the pod of sambac is subjected to a high-temperature sterilization process to obtain a sterilized pod of sambac, and the strain is combined with the sterilized pod of sambac to enhance the fermentation of the sterilized pod of sambac using the strain; wherein the sterilized pod of sambac is directly subjected to a dehydration process, a strain fermentation process and a sterilization process to obtain an enhanced fermented pod of sambac, and the enhanced fermented pod of sambac contains at least one volatile substance and at least one vanillin.

In a preferred embodiment of the present invention, the strain may be selected from bacillus velezensis ZN-S10.

The preferred embodiment of this invention has a sequence, which can be selected from Seq.1.

In a preferred embodiment of the invention, the volatile substance forms an aroma compound (e.g., alcohols, ketones, phenols, aldehydes, esters, terpenes, hydrocarbons, or carboxylic acids).

In a preferred embodiment of the present invention, the volatile substances and their aromas are formed by fermentation of the blanched vanilla pods using microorganisms.

In a preferred embodiment of the present invention, the dehydrated stalks of *Cymbidium goeringii* are dehydrated at a predetermined temperature, wherein the predetermined temperature includes the ranges from 15°C to 30°C, 16°C to 30°C, 17°C to 30°C, 18°C to 30°C, 19°C to 30°C, 20°C to 30°C, 21°C to 30°C, 22°C to 30°C, 23°C to 30°C, 24°C to 30°C, 25°C to 30°C, 26°C to 30°C, 27°C to 30°C, 28°C to 30°C, 29°C to 30°C, or other suitable temperature ranges.

In a preferred embodiment of the present invention, the sterilized stalks of *Cymbidium goeringii* undergo microbial fermentation catalysis at a predetermined temperature using a solar lamp catalytic unit. The predetermined temperature includes ranges from 15°C to 30°C, 16°C to 30°C, 17°C to 30°C, 18°C to 30°C, 19°C to 30°C, 20°C to 30°C, 21°C to 30°C, 22°C to 30°C, 23°C to 30°C, 24°C to 30°C, 25°C to 30°C, 26°C to 30°C, 27°C to 30°C, 28°C to 30°C, 29°C to 30°C, or other suitable temperature ranges.

In a preferred embodiment of the invention, the dehydrated and fermented vanilla pods are high-temperature dried at a predetermined temperature, which ranges from 30°C to 100°C.

S1: Steps

S10: Steps

S2: Steps

S3: Steps

S31: Steps

S32: Steps

S4: Steps

1: Unripe orchid pods

10: Mature orchid pods

10a: Dehydrated orchid pods

11: Enhanced fermentation of orchid pods

12: Dried and fermented fragrant pods (orchid pods)

100: Microbial strains

2: Baking Oven Unit

20: Refrigerated Drying Box Unit

21: Refrigerated Catalytic Converter Unit

200: Computer Unit

Figure 1: A block diagram illustrating a vanilla pod product produced by promoting fermentation with bacillus velezensis to form volatile substances, according to a first preferred embodiment of the present invention.

Figure 2: Flowchart of a first preferred embodiment of the present invention, illustrating a method for promoting the fermentation of volatile substances in vanilla pods using bacillus velezensis .

Figure 3: A block diagram of a vanilla pod product produced by promoting fermentation with bacillus velezensis to form volatile substances, according to a second preferred embodiment of the present invention.

Figure 4: Flowchart of a second preferred embodiment of the present invention, illustrating a method for promoting the fermentation of volatile substances in vanilla pods using bacillus velezensis .

Figure 5: A schematic diagram of the gas chromatography-mass spectra of volatile substances and vanillin in a scavenging product obtained by a method of promoting the fermentation of volatile substances in scavenging pods with bacillus velezensis according to a preferred embodiment of the present invention.

To provide a full understanding of the invention, preferred embodiments will be described in detail below with reference to the accompanying drawings, but these are not intended to limit the scope of the invention.

The preferred embodiment of the present invention, which uses bacillus velezensis to promote the fermentation of volatile substances in scavenging pods, its products and materials, is applicable to various scavenging pod varieties and their fermentation methods and systems. Furthermore, the preferred embodiment of the scavenging pod fermentation method and system can be selectively applied to various automated fermentation equipment, semi-automated fermentation equipment, or non-automated fermentation equipment, but it is not intended to limit the scope of application of the present invention.

The preferred embodiment of this invention, which uses bacillus velezensis to promote the fermentation of volatile substances in senna pods, and the products and materials thereof, are suitable for use in various food industries (e.g., seasonings for ice cream, cakes, cookies, chocolate, candy, coffee, tea, cola, and alcoholic beverages), various cosmetic industries (e.g., adding high-end flavorings, high-end essences, or high-end fragrances), or other related product industries, but is not intended to limit the scope of application of this invention.

The preferred embodiment of this invention, which uses bacillus velezensis to promote the fermentation of volatile substances in senna pods, and the products and materials thereof are suitable for various regional varieties, such as tropical and subtropical American and Caribbean varieties, African varieties, Madagascar varieties, Indian Peninsula varieties, or Southeast Asian varieties, but is not intended to limit the scope of application of this invention.

Figure 1 is a block diagram illustrating a vanilla pod product produced by promoting enhanced fermentation with bacillus velezensis to form volatile substances, according to a first preferred embodiment of the present invention. Referring to Figure 1, for example, the vanilla pod product produced by promoting enhanced fermentation with bacillus velezensis to form volatile substances according to a first preferred embodiment of the present invention uses a mature vanilla pod 10, which is appropriately obtained from an immature vanilla pod 1 (e.g., a vanilla cultivar of vanilla planifolia ).

Please refer again to Figure 1. For example, in the first preferred embodiment of the present invention, the pod product of bacillus velezensis promoting fermentation to form volatile substances adopts a microbial fermentation system. The microbial fermentation system may include a refrigerated drying unit 20, a drying unit (not shown) or a device with similar refrigerated drying or drying functions. The microbial fermentation system may also be connected to at least a computer unit 200 or a device with similar computer functions to achieve the purpose of providing a standard process for industrial production.

Referring again to Figure 1, for example, the computer unit 200 or a device with similar computer functions may be selected from a workstation computer, a desktop computer, a notebook or laptop computer, a tablet personal computer, a mobile communication device, a smartphone, or other devices with computer functions, but this is not intended to limit the scope of the invention.

Figure 2 illustrates a flowchart of a method for promoting the enrichment fermentation of volatile substances in sago palm pods using bacillus velezensis , according to a first preferred embodiment of the present invention. This corresponds to the sago palm pod product showing enrichment fermentation to form volatile substances, as shown in Figure 1. Referring to Figures 1 and 2, the enrichment fermentation method for volatile substances in sago palm pods according to a first preferred embodiment of the present invention includes step S10: First, for example, at least one strain 100 is appropriately prepared using suitable technical means (e.g., automated, semi-automated, or manual methods), and this strain 100 is selected from an enrichment fermentation strain (e.g., bacillus velezensis strain).

Please refer to Figures 1 and 2 again. For example, the strain can be selected from bacillus velezensis , bacillus velezensis ZN-S10 (which can be obtained from the "Active Natural Products and Biotechnology Service Center" of National Pingtung University of Science and Technology), or other strains similar to bacillus velezensis . The strain has a predetermined sequence, and the predetermined sequence can be selected from Seq.1 or a sequence similar to Seq.1.

Please refer to Figures 1 and 2 again. The first preferred embodiment of the present invention, the method for promoting the fermentation of volatile substances in sago palm pods by bacillus velezensis , includes step S1: Next, for example, the mature sago palm pod 10 is subjected to a high-temperature blanching operation by appropriate technical means (e.g., automated, semi-automated or manual) to obtain a blanched sago palm pod.

Please refer again to Figures 1 and 2. For example, the high-temperature sterilization operation is performed in a high-temperature steam unit, an insulated container unit, or a device with similar high-temperature steam or insulation functions. The high-temperature sterilization operation is carried out at a predetermined temperature, which includes 40°C to 100°C, 95°C to 100°C, or any temperature range therebetween.

Please refer again to Figures 1 and 2. The first preferred embodiment of the present invention, the method for promoting the enhanced fermentation of volatile substances in sago palm pods using bacillus velezensis , includes step S2: Next, for example, the strain 100 is appropriately combined with the bacillus velezensis in the mature sago palm pod 10 using appropriate technical means (e.g., automated, semi-automated or manual methods) (e.g., throwing, spraying, mechanical stirring or any combination thereof) so as to appropriately carry out enhanced fermentation of the mature sago palm pod 10 in the bacillus velezensis.

Please refer again to Figures 1 and 2, which illustrate the first preferred embodiment of this invention using bacillus. The velezensis method for promoting the enrichment of volatile substances in sago palm pods includes step S3: Next, for example, the mature sago palm pods 10 are subjected to a dehydration process (e.g., a moisture content of about 35% or other suitable moisture content after dehydration), a microbial fermentation process (e.g., a fermentation temperature of about 30°C or other suitable fermentation temperature), and a sterilization process by appropriate technical means (e.g., an automated, semi-automated, or manual method) to obtain an enriched fermented sago palm pod 11, which contains at least one volatile substance and at least one vanillin.

Please refer again to Figures 1 and 2. For example, the volatile substance forms an aroma compound (e.g., alcohols, ketones, phenols, aldehydes, esters, terpenes, hydrocarbons, or carboxylic acids), and the mature, blanched pods of the orchid 10 are fermented using strain 100 to adjust and form the volatile substance and its odor.

Please refer again to Figures 1 and 2. For example, any stage of the dehydration, fermentation, or sterilization process can be performed in the refrigerated drying unit 20 or a device with similar refrigerated drying or drying functions to achieve the goals of reducing manufacturing costs, reducing process energy consumption, simplifying the overall process, and reducing manpower.

Referring again to Figures 1 and 2, for example, in another preferred embodiment of the invention, the entire series of operations—dehydration, microbial fermentation, and sterilization—are performed within the refrigerated drying unit 20 or a device with similar refrigerated drying or drying functions. The mature, blanched orchid pods 10 are processed entirely in one step within the refrigerated drying unit 20 or a device with similar refrigerated drying or drying functions, thereby achieving the goals of reducing manufacturing costs, reducing process energy consumption, simplifying the overall process, and reducing manpower.

Please refer again to Figures 1 and 2. For example, in another preferred embodiment of the invention, all the dehydration, fermentation, and sterilization processes are performed in the refrigerated drying unit 20 or a device with similar refrigerated drying or drying functions. This simplifies the overall process, reduces the number of times the pods are moved (moved in and out), and avoids continuous changes in operating conditions under high temperature, outdoor sunlight, and cooling. Furthermore, avoiding outdoor sunlight and cooling of the pods reduces contamination from external dust or insect (dead) environments.

Please refer again to Figures 1 and 2. For example, the mature pods of *Cymbidium goeringii* 10, after being dehydrated, are dehydrated (gradually and slowly) at a predetermined temperature to prevent the growth of mold or other microorganisms. This predetermined temperature ranges from 15°C to 30°C, 16°C to 30°C, 17°C to 30°C, 18°C to 30°C, and 19°C. Between 20°C and 30°C, 21°C and 30°C, 22°C and 30°C, 23°C and 30°C, 24°C and 30°C, 25°C and 30°C, 26°C and 30°C, 27°C and 30°C, 28°C and 30°C, 29°C and 30°C, or other suitable temperature ranges.

Please refer again to Figures 1 and 2. For example, the mature orchid pods 10, which have been sterilized, are subjected to microbial fermentation at a predetermined temperature for a predetermined time (e.g., 7 days or other catalytic periods) using a solar lamp or a device with a similar sterilization function. The solar lamp is located in the refrigerated drying unit 20 or a device with a similar refrigerated drying or drying function, and the predetermined temperature includes 15°C to 30°C, with 16°C being an additional temperature. Between 17°C and 30°C, 18°C and 30°C, 19°C and 30°C, 20°C and 30°C, 21°C and 30°C, 22°C and 30°C, 23°C and 30°C, 24°C and 30°C, 25°C and 30°C, 26°C and 30°C, 27°C and 30°C, 28°C and 30°C, 29°C and 30°C, or other suitable temperature ranges.

Please refer again to Figures 1 and 2. For example, if the predetermined temperature is limited to between 15°C and 30°C, the growth of microorganisms or other microorganisms in the sago palm pods and their contamination can be inhibited throughout the entire process, thereby improving the quality of the fermented product with increased volatile substances from the sago palm pods.

Please refer again to Figures 1 and 2. The first preferred embodiment of the present invention, the method for promoting the enrichment of volatile substances in sago palm pods using bacillus velezensis , includes step S4: Next, for example, the enriched sago palm pod 11 is appropriately subjected to a high-temperature drying operation using appropriate technical means (e.g., automated, semi-automated or manual) to obtain a dried enriched sago palm pod 12.

Please refer again to Figures 1 and 2. For example, the dried, enriched fermented stalk of orchid 12 is produced from an orchid pod using the enriched fermentation method for volatile substances in orchid pods according to the first preferred embodiment of the present invention. The dried, enriched fermented stalk of orchid 12 is rich in at least one volatile substance, at least one vanillin, and other active ingredients.

Please refer again to Figures 1 and 2. For example, the high-temperature drying operation is performed in an oven unit 2 or a device with similar baking functions (e.g., a hot air circulation convection device), and the fermented stalks 11 are dried at a predetermined temperature, which includes 30°C to 100°C.

Figure 3 is a block diagram illustrating a scavenging product of bacillus velezensis promoting fermentation to form volatile substances, according to a second preferred embodiment of the present invention. Referring to Figure 3, for example, the scavenging product of bacillus velezensis promoting fermentation to form volatile substances in the second preferred embodiment of the present invention employs a microbial fermentation system. This microbial fermentation system may optionally include a refrigerated drying unit 20, a refrigerated catalytic unit 21, or a device with similar refrigerated drying or refrigerated catalytic functions. Furthermore, the microbial fermentation system may optionally be connected to at least one computer unit 200 or a device with similar computer functions to achieve the purpose of providing a standard industrial production process.

Figure 4 illustrates a flowchart of a second preferred embodiment of the present invention, showing a method for promoting the enrichment of volatile substances in sago palm pods using bacillus velezensis . This corresponds to the sago palm pod product shown in Figure 3, which uses bacillus velezensis to promote enrichment fermentation and form volatile substances. Referring to Figures 3 and 4, the method for promoting the enrichment of volatile substances in sago palm pods using bacillus velezensis in the second preferred embodiment of the present invention includes step S10: First, for example, at least one strain 100 is appropriately prepared using suitable technical means (e.g., automated, semi-automated, or manual methods), and the strain 100 is selected from an enrichment fermentation strain.

Please refer again to Figures 3 and 4. The second preferred embodiment of the present invention, the method for promoting the fermentation of volatile substances in sago palm pods by bacillus velezensis , includes step S1: Next, for example, the mature sago palm pod 10 is subjected to a high-temperature blanching operation by appropriate technical means (e.g., automated, semi-automated or manual) to obtain a blanched sago palm pod.

Please refer again to Figures 3 and 4. The second preferred embodiment of the present invention, the method for promoting the enrichment of volatile substances in sago palm pods using bacillus velezensis , includes step S2: Next, for example, the strain 100 is appropriately combined with the blanched sago palm pods of the mature sago palm pods 10 by appropriate technical means (e.g., automated, semi-automated or manual methods) so as to appropriately carry out enrichment fermentation of the blanched sago palm pods of the mature sago palm pods 10 using the strain 100.

Please refer again to Figures 3 and 4. The second preferred embodiment of the present invention, the method for promoting the increase of volatile substances in sago palm pods by bacillus velezensis , includes step S31: Next, for example, the mature sago palm pod 10 is directly subjected to a dehydration operation by appropriate technical means (e.g., automated, semi-automated or manual) to obtain a dehydrated sago palm pod 10a.

Please refer again to Figures 3 and 4. For example, the dehydration operation is performed in the refrigerated drying unit 20 or a device with similar refrigerated drying or drying functions.

Please refer again to Figures 3 and 4. The second preferred embodiment of the present invention, the method for promoting the enrichment of volatile substances in sago palm pods using bacillus velezensis , includes step S32: Next, for example, the dehydrated sago palm pod 10a is appropriately subjected to a microbial fermentation operation and a sterilization operation by appropriate technical means (e.g., automated, semi-automated or manual) to obtain an enriched fermented sago palm pod 11.

Please refer again to Figures 3 and 4. For example, the fermentation process is carried out in the refrigerated catalytic unit 21 or a device with similar refrigerated or room-temperature catalytic functions to avoid high-temperature catalytic fermentation and save processing energy.

Please refer again to Figures 3 and 4. The second preferred embodiment of the present invention, the method for promoting the enrichment of volatile substances in sago palm pods with bacillus velezensis , includes step S4: Next, for example, the enriched sago palm pod 11 is appropriately subjected to a high-temperature drying operation by appropriate technical means (e.g., automated, semi-automated or manual) to obtain a dried enriched sago palm pod 12.

Please refer again to Figures 3 and 4. For example, the high-temperature drying operation is performed in an oven unit 2 or a device with similar baking functions (e.g., a hot air circulation convection device), and the dehydrated and fermented stalks 11 are dried at a predetermined temperature, which includes 30°C to 100°C.

Figure 5 shows a schematic diagram of the gas chromatography-mass spectra of volatile substances and vanillin in a scavenging product obtained by a bacillus velezensis -promoted fermentation method for enhancing the fermentation of volatile substances in scavenging pods, according to a preferred embodiment of the present invention. Please refer to Figure 5. For example, the vanilla pod product of the preferred embodiment of the present invention, which produces volatile substances through fermentation, was analyzed by gas chromatography-mass spectrometry (SPME-GCMS) to obtain several volatile substances (as shown on the left side of Figure 5) and oxalaldehyde (as shown in the middle of Figure 5, which is the catalytic conversion of vanillin from vanillin alcohol).

Referring again to Figure 5, for example, the volatile substances in the vanilla pod product produced by the enhanced fermentation of the preferred embodiment of the present invention may include alcohol compounds (e.g., 1-Octen-3-ol, 2-ethyl-1-Hexanol, 2-propyl-1-Heptanol, 4,8-dimethyl1-Nonanol, 3-Hexanol, 3,5-dimethyl, 1-Octanol, 2-butyl, Phenol, 2-methoxy-4-methyl, Isotridecanol, or other alcohol compounds), but this is not intended to limit the scope of the present invention.

Referring again to Figure 5, for example, the volatile substances in the vanilla pod product of the preferred embodiment of the present invention, which enhances fermentation to form volatile substances, may include ester compounds (e.g., pentafluoropropionic acid or other compounds), but this is not intended to limit the scope of the present invention.

Referring again to the left side of Figure 5, for example, the volatile substances in the vanilla pod product produced by the enhanced fermentation of volatile substances in the preferred embodiment of the present invention may include aldehyde compounds (e.g., 2-Heptenal, Octanal, Nonanal, Decanal, Vanillin, or other aldehyde compounds), but this is not intended to limit the scope of the present invention.

Referring again to Figure 5, for example, the volatile substances in the pod-based product of the preferred embodiment of the present invention, which enhances fermentation to form volatile substances, may include ketone compounds (e.g., 2-Pentanone, 4-hydroxy, or other ketone compounds), but this is not intended to limit the scope of the invention.

Please refer again to Figure 5. For example, the volatile substances in the vanilla pod product of the preferred embodiment of the invention that enhances fermentation to form volatile substances may include hydrocarbon compounds (e.g., 2,4-Dimethyl-1-heptene, Styrene, 1-Octene, 3,7-dimethyl, Limonene, Copaene, Bicyclo[3.1.1]hept-2-ene, 2,6-dimethyl-6-(4-methyl-3-pentenyl)). The following compounds are permitted: cederne, isooctane (ethenyloxy), Decane (4-methyl), Cyclohexane (isothiocyanato), Dodecane (2,6,10-trimethyl), Cyclopropane (1,1-dimethyl-2-(1-methylethoxy)-3-(3-methyl-1-pentynyl), or other hydrocarbon compounds; however, they are not intended to limit the scope of this invention.

Referring again to Figure 5, for example, the volatile substances in the pod-based product of the preferred embodiment of the present invention, which enhances fermentation to form volatile substances, may include carboxylic acid compounds (e.g., sulfurous acid, trifluoroacetic acid, heptafluorobutyric acid, or other carboxylic acid compounds), but this is not intended to limit the scope of the present invention.

The above experimental data are preliminary results obtained under specific conditions and are intended only to facilitate understanding or reference of the technical content of this invention. Further related experiments are required. This experimental data and its results are not intended to limit the scope of the claims of this invention.

The foregoing preferred embodiments are merely illustrative of the invention and its technical features. The technology of these embodiments can still be implemented with various substantially equivalent modifications and/or substitutions; therefore, the scope of the invention shall be determined by the scope defined in the appended patent application. This copyright application is limited to the purposes for which a patent application in the Republic of China is filed.

1: Unripe orchid pods

10: Mature orchid pods

11: Enhanced fermentation of orchid pods

12: Dried and fermented fragrant pods (orchid pods)

100: Microbial strains

2: Baking Oven Unit

20: Refrigerated Drying Box Unit

200: Computer Unit

Claims (10)

A method for promoting the enrichment fermentation of volatile substances in sago palm pods using bacillus velezensis , comprising: preparing at least one strain, wherein the strain is selected from an enrichment fermentation strain bacillus velezensis. The invention relates to velezensis and related strains; to subject a pod of sambac to a high-temperature sterilization process to obtain a sterilized pod of sambac; to combine the strain with the sterilized pod of sambac to enhance the fermentation of the sterilized pod of sambac; and to directly subject the sterilized pod of sambac to a dehydration process, a fermentation process, and a sterilization process to obtain an enhanced fermented pod of sambac, wherein the enhanced fermented pod of sambac contains at least one volatile substance and at least one vanillin. According to the method for promoting the fermentation of volatile substances in sago palm pods by using bacillus velezensis as described in claim 1, the strain may be selected from bacillus velezensis ZN-S10. According to the method for promoting the fermentation of volatile substances in stalks of sambac using bacillus velezensis as described in claim 1, wherein the strain has a sequence that can be selected from Seq.1. The method for promoting the fermentation of volatile substances in stalks of stalks of stalks by using bacillus velezensis as described in claim 1 of the patent application, wherein the volatile substances form an aroma substance. According to the first claim of the patent application, the method for promoting the fermentation of volatile substances in sago palm pods by using bacillus velezensis is described, wherein the blanched sago palm pods are fermented by the strain to adjust and form the volatile substances and their aroma. A pod product from pods that utilizes bacillus velezensis to promote enhanced fermentation and form volatile substances comprises: at least one pod, wherein the pod is subjected to a high-temperature blanching process to obtain a blanched pod, and at least one microbial strain is incorporated into the blanched pod to enhance fermentation of the blanched pod using the microbial strain, wherein the microbial strain is selected from a bacillus velezensis-based enhanced fermentation strain. The velezensis and related strains; wherein the sterilized velezensis pods are directly subjected to a dehydration process, a strain fermentation process and a sterilization process to obtain an enhanced fermented velezensis pod, and the enhanced fermented velezensis pod contains at least one volatile substance and at least one vanillin. According to claim 6, the sago palm pod product that uses bacillus velezensis to promote fermentation and form volatile substances, wherein the strain can be selected from bacillus velezensis ZN-S10. According to claim 6, the sago palm pod product that uses bacillus velezensis to promote fermentation and form volatile substances, wherein the strain has a sequence that can be selected from Seq.1. According to claim 6, the sago palm pod product that uses bacillus velezensis to promote fermentation and form volatile substances, wherein the volatile substances form a fragrance substance. According to claim 6, the sago palm pod product that uses bacillus velezensis to promote fermentation and form volatile substances, wherein the bacillus velezensis pod is fermented by the strain to form the volatile substances and their aroma.