CN105803005A - Method for promoting synthesis of squalene from schizochytrium limacinum by virtue of exogenous regulatory factor - Google Patents
Method for promoting synthesis of squalene from schizochytrium limacinum by virtue of exogenous regulatory factor Download PDFInfo
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Abstract
本发明公开了一种外源调控因子促进裂殖壶菌合成角鲨烯的方法，包括如下步骤：在利用裂殖壶菌发酵合成角鲨烯时，在发酵前或发酵中向发酵培养基中添加一定浓度的外源调控因子，以促进角鲨烯的合成，其中，所述外源调控因子为醇。本发明通过简单有效的发酵调控，提高裂殖壶菌中角鲨烯含量，效果明显，可应用于大规模生产。且该方法简单方便，具有经济效益。
The invention discloses a method for exogenous regulatory factors to promote the synthesis of squalene by Schizochytrium fungus, which comprises the following steps: when using Schizochytrium fungus to ferment and synthesize squalene, adding squalene to the fermentation medium before or during fermentation A certain concentration of exogenous regulatory factor is added to promote the synthesis of squalene, wherein the exogenous regulatory factor is alcohol. The invention improves the content of squalene in the Schizochytrium through simple and effective fermentation regulation, the effect is obvious, and it can be applied to large-scale production. Moreover, the method is simple and convenient, and has economic benefits.
Description
技术领域technical field
本发明属于生物技术领域，特别涉及一种外源调控因子促进裂殖壶菌合成角鲨烯的方法。The invention belongs to the field of biotechnology, and in particular relates to a method for promoting the synthesis of squalene by Schizochytrium fungus by exogenous regulatory factors.
背景技术Background technique
裂殖壶菌(Schizochytrium或Aurantiochytrium)是一种海洋真菌，其菌体内积累有大量的DHA，所以它是目前生产DHA的重要菌株。除了DHA之外，裂殖壶菌菌体中也含有角鲨烯等物质。角鲨烯是一种脂质不皂化物，具有增强机体免疫能力、抗衰老、抗疲劳、抗肿瘤等多种生理功能，是一种具有防病治病作用的生物活性物质，并广泛应用于医药、化妆品等领域。目前角鲨烯的主要来源是鲨鱼肝油，但渔业资源的枯竭促使人们寻找其他的角鲨烯来源。利用微生物发酵法来生产角鲨烯是一种很有潜力的角鲨烯来源。Schizochytrium (Schizochytrium or Aurantiochytrium) is a kind of marine fungus, which accumulates a large amount of DHA in its body, so it is an important strain for producing DHA at present. In addition to DHA, Schizochytrium also contains substances such as squalene. Squalene is a lipid unsaponifiable substance, which has various physiological functions such as enhancing the body's immunity, anti-aging, anti-fatigue, and anti-tumor. It is a biologically active substance with the effect of preventing and curing diseases, and is widely used medicine, cosmetics and other fields. Currently the main source of squalene is shark liver oil, but the depletion of fishery resources has prompted people to look for other sources of squalene. The use of microbial fermentation to produce squalene is a potential source of squalene.
发明内容Contents of the invention
本发明的一个目的是解决至少上述问题和/或缺陷，并提供至少后面将说明的优点。An object of the present invention is to solve at least the above problems and/or disadvantages and to provide at least the advantages as will be described hereinafter.
针对现有技术中利用裂殖壶菌进行发酵生产角鲨烯含量低的特点，本发明还有一个目的是提供一种外源调控因子促进裂殖壶菌合成角鲨烯的方法，本发明的方法通过抑制裂殖壶菌合成角鲨烯途径的竞争途径，使更多的前体物质和还原剂流向角鲨烯合成，以提高菌体中的角鲨烯含量。In view of the low content of squalene produced by fermentation by Schizochytrium in the prior art, another object of the present invention is to provide a method for exogenous regulatory factors to promote the synthesis of squalene by Schizochytrium. Methods By inhibiting the competitive pathway of the synthesis of squalene in Schizochytrium, more precursors and reducing agents flow to the synthesis of squalene, so as to increase the content of squalene in the bacteria.
本发明提供的技术方案为：The technical scheme provided by the invention is:
一种外源调控因子促进裂殖壶菌合成角鲨烯的方法，包括如下步骤：A method for exogenous regulatory factors to promote the synthesis of squalene by Schizochytrium, comprising the steps of:
在利用裂殖壶菌发酵合成角鲨烯时，在发酵前或发酵中向发酵培养基中添加一定浓度的外源调控因子，以促进角鲨烯的合成，其中，所述外源调控因子为醇。When using Schizochytrium to ferment and synthesize squalene, add a certain concentration of exogenous regulatory factors to the fermentation medium before or during fermentation to promote the synthesis of squalene, wherein the exogenous regulatory factors are alcohol.
优选的是，所述的外源调控因子促进裂殖壶菌合成角鲨烯的方法中，所述醇为甲醇、乙醇、丙醇、丁醇、异丙醇、异丁醇、异戊醇和巴豆醇中的任意一种或几种。Preferably, in the method that described exogenous regulation factor promotes Schizochytrium to synthesize squalene, described alcohol is methanol, ethanol, propanol, butanol, isopropanol, isobutanol, isoamyl alcohol and croton Any one or more of alcohols.
优选的是，所述的外源调控因子促进裂殖壶菌合成角鲨烯的方法中，所述醇为甲醇、乙醇、丙醇、丁醇、异丙醇、异丁醇、异戊醇和巴豆醇中的任意一种，Preferably, in the method that described exogenous regulation factor promotes Schizochytrium to synthesize squalene, described alcohol is methanol, ethanol, propanol, butanol, isopropanol, isobutanol, isoamyl alcohol and croton any of the alcohols,
当所述醇为甲醇时，所述一定浓度为40-100g/L，When the alcohol is methanol, the certain concentration is 40-100g/L,
当所述醇为乙醇时，所述一定浓度为20-50g/L，When the alcohol is ethanol, the certain concentration is 20-50g/L,
当所述醇为丙醇时，所述一定浓度为8-20g/L，When the alcohol is propanol, the certain concentration is 8-20g/L,
当所述醇为丁醇时，所述一定浓度为4-10g/L，When the alcohol is butanol, the certain concentration is 4-10g/L,
当所述醇为异丙醇时，所述一定浓度为20-50g/L，When the alcohol is isopropanol, the certain concentration is 20-50g/L,
当所述醇为异丁醇时，所述一定浓度为4-10g/L，When the alcohol is isobutanol, the certain concentration is 4-10g/L,
当所述醇为异戊醇时，所述一定浓度为2-5g/L，When the alcohol is isoamyl alcohol, the certain concentration is 2-5g/L,
当所述醇为巴豆醇时，所述一定浓度为4-10g/L。When the alcohol is crotyl alcohol, the certain concentration is 4-10 g/L.
优选的是，所述的外源调控因子促进裂殖壶菌合成角鲨烯的方法中，当所述醇为甲醇时，所述一定浓度为80g/L，Preferably, in the method for promoting the synthesis of squalene by Schizochytrium by exogenous regulatory factors, when the alcohol is methanol, the certain concentration is 80g/L,
当所述醇为乙醇时，所述一定浓度为40g/L，When the alcohol is ethanol, the certain concentration is 40g/L,
当所述醇为丙醇时，所述一定浓度为16g/L，When the alcohol is propanol, the certain concentration is 16g/L,
当所述醇为丁醇时，所述一定浓度为8g/L，When the alcohol is butanol, the certain concentration is 8g/L,
当所述醇为异丙醇时，所述一定浓度为40g/L，When the alcohol is Virahol, the certain concentration is 40g/L,
当所述醇为异丁醇时，所述一定浓度为8g/L，When the alcohol is isobutanol, the certain concentration is 8g/L,
当所述醇为异戊醇时，所述一定浓度为4g/L，When the alcohol is isoamyl alcohol, the certain concentration is 4g/L,
当所述醇为巴豆醇时，所述一定浓度为8g/L。When the alcohol is crotyl alcohol, the certain concentration is 8g/L.
优选的是，所述的外源调控因子促进裂殖壶菌合成角鲨烯的方法中，向所述发酵培养基中添加所述外源调控因子的方式为一次性加入、分次加入或者持续流动加入。Preferably, in the method for promoting the synthesis of squalene by Schizochytrium by the exogenous regulatory factor, the method of adding the exogenous regulatory factor to the fermentation medium is one-time addition, divided addition or continuous addition. Flow joins.
本发明至少包括以下有益效果：The present invention at least includes the following beneficial effects:
本发明通过添加外源调控因子，抑制合成DHA的多不饱和脂肪酸合成酶途径，实现代谢向甲羟戊酸途径的调控，从而促进角鲨烯的合成，提高裂殖壶菌菌体中角鲨烯含量。依照本发明的方法可显著提裂殖壶菌菌体中高角鲨烯的含量；并且本发明操作简单，不需要额外的人工和设备，仅通过较低的投入，就可利用现有的裂殖壶菌发酵技术实现角鲨烯的生产。The present invention inhibits the polyunsaturated fatty acid synthase pathway of synthesizing DHA by adding exogenous regulatory factors, and realizes the regulation of metabolism to the mevalonate pathway, thereby promoting the synthesis of squalene and increasing the content of squalene in Schizochytrium thallus. ene content. According to the method of the present invention, the content of high squalene in the Schizochytrium thalline can be significantly improved; and the present invention is simple to operate, does not require additional labor and equipment, and can utilize the existing fission Chytrium fermentation technology realizes the production of squalene.
本发明的其它优点、目标和特征将部分通过下面的说明体现，部分还将通过对本发明的研究和实践而为本领域的技术人员所理解。Other advantages, objectives and features of the present invention will partly be embodied through the following descriptions, and partly will be understood by those skilled in the art through the study and practice of the present invention.
附图说明Description of drawings
图1为本发明其中一个实施例中角鲨烯含量测定结果图。Fig. 1 is a graph showing the determination results of squalene content in one of the embodiments of the present invention.
具体实施方式detailed description
下面结合附图对本发明做进一步的详细说明，以令本领域技术人员参照说明书文字能够据以实施。The present invention will be further described in detail below in conjunction with the accompanying drawings, so that those skilled in the art can implement it with reference to the description.
应当理解，本文所使用的诸如“具有”、“包含”以及“包括”术语并不配出一个或多个其它元件或其组合的存在或添加。It should be understood that terms such as "having", "comprising" and "including" as used herein do not entail the presence or addition of one or more other elements or combinations thereof.
本发明提供一种外源调控因子促进裂殖壶菌合成角鲨烯的方法，包括如下步骤：The invention provides a method for exogenous regulatory factors to promote the synthesis of squalene by Schizochytrium, comprising the following steps:
在利用裂殖壶菌发酵合成角鲨烯时，在发酵前或发酵中向发酵培养基中添加一定浓度的外源调控因子，以促进角鲨烯的合成，其中，所述外源调控因子为醇。When using Schizochytrium to ferment and synthesize squalene, add a certain concentration of exogenous regulatory factors to the fermentation medium before or during fermentation to promote the synthesis of squalene, wherein the exogenous regulatory factors are alcohol.
在裂殖壶菌胞内，DHA作为一种多不饱和脂肪酸，可以增强细胞膜的流动性和通透性，它的合成途径是多不饱和脂肪酸合成酶途径合成。角鲨烯则是由甲羟戊酸途径合成。这两种途径使用一样的前体(乙酰辅酶A)和一样的还原剂(NADPH)，所以它们互为竞争途径。当在培养基中加入外源调控因子如丁醇等时，这些外源调控因子可通过细胞膜渗入裂殖壶菌细胞，并对细胞产生毒害作用。作为应激反应，裂殖壶菌细胞会抑制多不饱和脂肪酸合成酶途径并减少DHA等多不饱和脂肪酸的合成，使得细胞膜通透性下降，外源调控因子的渗入减少，细胞毒害作用减弱；同时作为多不饱和脂肪酸合成酶途径的竞争途径，甲羟戊酸途径由于得到更多的前体和还原剂而被增强，因而可以促进角鲨烯的积累。In Schizochytrium cells, DHA, as a polyunsaturated fatty acid, can enhance the fluidity and permeability of the cell membrane, and its synthesis pathway is polyunsaturated fatty acid synthase pathway. Squalene is synthesized by the mevalonate pathway. Both pathways use the same precursor (acetyl-CoA) and the same reducing agent (NADPH), so they are competing pathways. When exogenous regulatory factors such as butanol are added to the medium, these exogenous regulatory factors can penetrate into the cells of Schizochytrium through the cell membrane and produce toxic effects on the cells. As a stress response, Schizochytrium cells will inhibit the polyunsaturated fatty acid synthase pathway and reduce the synthesis of polyunsaturated fatty acids such as DHA, which will reduce the permeability of the cell membrane, reduce the infiltration of exogenous regulatory factors, and weaken the cytotoxic effect; At the same time, as a competitive pathway of the polyunsaturated fatty acid synthase pathway, the mevalonate pathway is enhanced by getting more precursors and reducing agents, which can promote the accumulation of squalene.
依照本发明的方法可显著提裂殖壶菌菌体中高角鲨烯的含量；并且本发明操作简单，不需要额外的人工和设备，仅通过较低的投入，就可利用现有的裂殖壶菌发酵技术实现角鲨烯的大量生产。According to the method of the present invention, the content of high squalene in the Schizochytrium thalline can be significantly improved; and the present invention is simple to operate, does not require additional labor and equipment, and can utilize the existing fission Chytrium fermentation technology realizes mass production of squalene.
在本发明的一些实施例中，作为优选，所述醇为甲醇、乙醇、丙醇、丁醇、异丙醇、异丁醇、异戊醇和巴豆醇中的任意一种或几种。In some embodiments of the present invention, preferably, the alcohol is any one or more of methanol, ethanol, propanol, butanol, isopropanol, isobutanol, isoamyl alcohol and crotyl alcohol.
在上述方案中，所述醇为甲醇、乙醇、丙醇、丁醇、异丙醇、异丁醇、异戊醇和巴豆醇中的任意一种时，作为优选，In the above scheme, when the alcohol is any one of methanol, ethanol, propanol, butanol, isopropanol, isobutanol, isoamyl alcohol and crotyl alcohol, as preferably,
当所述醇为甲醇时，所述一定浓度为40-100g/L，When the alcohol is methanol, the certain concentration is 40-100g/L,
当所述醇为乙醇时，所述一定浓度为20-50g/L，When the alcohol is ethanol, the certain concentration is 20-50g/L,
当所述醇为丙醇时，所述一定浓度为8-20g/L，When the alcohol is propanol, the certain concentration is 8-20g/L,
当所述醇为丁醇时，所述一定浓度为4-10g/L，When the alcohol is butanol, the certain concentration is 4-10g/L,
当所述醇为异丙醇时，所述一定浓度为20-50g/L，When the alcohol is isopropanol, the certain concentration is 20-50g/L,
当所述醇为异丁醇时，所述一定浓度为4-10g/L，When the alcohol is isobutanol, the certain concentration is 4-10g/L,
当所述醇为异戊醇时，所述一定浓度为2-5g/L，When the alcohol is isoamyl alcohol, the certain concentration is 2-5g/L,
当所述醇为巴豆醇时，所述一定浓度为4-10g/L。When the alcohol is crotyl alcohol, the certain concentration is 4-10 g/L.
在上述方案中，作为优选，In the above scheme, as a preference,
当所述醇为甲醇时，所述一定浓度为80g/L，When the alcohol is methanol, the certain concentration is 80g/L,
当所述醇为乙醇时，所述一定浓度为40g/L，When the alcohol is ethanol, the certain concentration is 40g/L,
当所述醇为丙醇时，所述一定浓度为16g/L，When the alcohol is propanol, the certain concentration is 16g/L,
当所述醇为丁醇时，所述一定浓度为8g/L，When the alcohol is butanol, the certain concentration is 8g/L,
当所述醇为异丙醇时，所述一定浓度为40g/L，When the alcohol is Virahol, the certain concentration is 40g/L,
当所述醇为异丁醇时，所述一定浓度为8g/L，When the alcohol is isobutanol, the certain concentration is 8g/L,
当所述醇为异戊醇时，所述一定浓度为4g/L，When the alcohol is isoamyl alcohol, the certain concentration is 4g/L,
当所述醇为巴豆醇时，所述一定浓度为8g/L。When the alcohol is crotyl alcohol, the certain concentration is 8g/L.
在本发明的其中一些实施例中，作为优选，向所述发酵培养基中添加所述外源调控因子的方式为一次性加入、分次加入或者持续流动加入。In some embodiments of the present invention, preferably, the method of adding the exogenous regulatory factor to the fermentation medium is one-time addition, batch addition or continuous flow addition.
以下实施中，所使用的角鲨烯的测定方法为：In following implementation, the assay method of the squalene used is:
冷冻干燥：将发酵液收集在离心管中，离心去上清，用蒸馏水洗涤沉淀2次，再次离心；将沉淀置于冷冻离心机中冷冻干燥24-48小时，直到沉淀重量恒定；称取细胞干重的质量。Freeze-drying: Collect the fermentation broth in a centrifuge tube, centrifuge to remove the supernatant, wash the precipitate twice with distilled water, and centrifuge again; place the precipitate in a refrigerated centrifuge and freeze-dry for 24-48 hours until the weight of the precipitate is constant; weigh the cells dry weight mass.
超声破碎提取油脂：将少量菌体置和蒸馏水置于另一离心管中，漩涡振荡使之充分混匀，然后在冰浴条件下超声破碎；破碎条件为：开3秒，停3秒，循环100次；在离心管中加入正己烷，摇床振荡8小时左右；吸取离心管中上层的有机相，低温减压蒸馏出去溶剂，得到油脂；在测定角鲨烯之前，需要把样品甲酯化。Ultrasonic crushing to extract oil: put a small amount of bacteria and distilled water in another centrifuge tube, vortex to mix well, and then ultrasonically crush in an ice bath; the crushing conditions are: open for 3 seconds, stop for 3 seconds, and cycle 100 times; add n-hexane into the centrifuge tube, and shake the shaker for about 8 hours; absorb the organic phase in the upper layer of the centrifuge tube, and distill off the solvent under low temperature and reduced pressure to obtain oil; before measuring squalene, the sample needs to be methylated .
气相色谱法测定角鲨烯：日本岛津GC2010，FID检测器，Rtx-5色谱柱，进样口温度250度，检测器温度260度，柱温150度保持2分钟，然后以8度/分钟升温至250度，保持5分钟。Determination of squalene by gas chromatography: Shimadzu GC2010, FID detector, Rtx-5 chromatographic column, inlet temperature 250 degrees, detector temperature 260 degrees, column temperature 150 degrees for 2 minutes, and then at 8 degrees/minute Raise the temperature to 250 degrees and hold for 5 minutes.
实施例1Example 1
将裂殖壶菌B4D1接种到发酵培养基中，发酵培养基的组分为葡萄糖80g/L、酵母粉4g/L、大豆蛋白胨4g/L、海水晶15g/L，MgCl2·6H2O6.4g/L、CaCl2·2H2O1g/L、MgSO4·7H2O5g/L、KCl2g/L。在接种后24小时时，分别一次性加入外源调控因子甲醇、乙醇、丙醇、丁醇、巴豆醇。利用生物传感仪测定培养基中的葡萄糖浓度，当培养基中的葡萄糖浓度为0g/L时，停止发酵。Schizochytrium B4D1 was inoculated into the fermentation medium, and the components of the fermentation medium were glucose 80g/L, yeast powder 4g/L, soybean peptone 4g/L, sea crystal 15g/L, MgCl 2 6H 2 O6. 4g/L, CaCl 2 2H 2 O 1g/L, MgSO 4 7H 2 O 5g/L, KCl 2g/L. At 24 hours after inoculation, the exogenous regulatory factors methanol, ethanol, propanol, butanol and crotyl alcohol were added in one go. Utilize biosensor to measure the glucose concentration in the culture medium, when the glucose concentration in the culture medium is 0g/L, stop fermentation.
实验结果如表1所示：The experimental results are shown in Table 1:
其中细胞干重是指每升发酵液获得的菌体量，角鲨烯含量是指角鲨烯与菌体干重的比例。Wherein, the dry cell weight refers to the amount of cells obtained per liter of fermentation broth, and the squalene content refers to the ratio of squalene to the dry weight of cells.
由表1可以看出，对照组所获得的菌体中，角鲨烯的含量较低，但是在加入了甲醇、乙醇、丙醇、丁醇、巴豆醇等调控因子后，角鲨烯的含量显著升高。As can be seen from Table 1, in the thalline obtained by the control group, the content of squalene is low, but after adding regulatory factors such as methanol, ethanol, propanol, butanol, crotyl alcohol, the content of squalene Significantly increased.
表1Table 1
实施例2Example 2
将裂殖壶菌ATCC20888接种到发酵培养基中，发酵培养基的组分为葡萄糖80g/L、酵母粉4g/L、大豆蛋白胨4g/L、海水晶15g/L。在接种后36小时和50小时时，分2次加入外源调控因子甲醇、乙醇、丙醇、丁醇、巴豆醇，每次加入的浓度为最终外源调控因子浓度的一半。利用生物传感仪测定培养基中的葡萄糖浓度，当培养基中的葡萄糖浓度为0g/L时，停止发酵。Schizochytrium ATCC20888 was inoculated into the fermentation medium, and the components of the fermentation medium were glucose 80g/L, yeast powder 4g/L, soybean peptone 4g/L, and sea crystal 15g/L. At 36 hours and 50 hours after inoculation, the exogenous regulatory factors methanol, ethanol, propanol, butanol, and crotyl alcohol were added twice, and the concentration of each addition was half of the final exogenous regulatory factor concentration. Utilize biosensor to measure the glucose concentration in the culture medium, when the glucose concentration in the culture medium is 0g/L, stop fermentation.
实验结果如表2所示：The experimental results are shown in Table 2:
表2Table 2
其中细胞干重是指每升发酵液获得的菌体量，角鲨烯含量是指角鲨烯与菌体干重的比例。Wherein, the dry cell weight refers to the amount of cells obtained per liter of fermentation broth, and the squalene content refers to the ratio of squalene to the dry weight of cells.
裂殖壶菌ATCC20888对外源调控因子的耐受程度较弱，所以可以选择分次加入外源调控因子的策略。由表2可以看出，分次加入外源调控因子的策略同样可以实现促进角鲨烯积累的目的。Schizochytrium ATCC20888 has weak tolerance to exogenous regulatory factors, so the strategy of adding exogenous regulatory factors in stages can be chosen. It can be seen from Table 2 that the strategy of adding exogenous regulatory factors in stages can also achieve the purpose of promoting the accumulation of squalene.
实施例3Example 3
将裂殖壶菌B4D1接种到发酵培养基中，发酵培养基的组分为葡萄糖80g/L、酵母粉4g/L、大豆蛋白胨4g/L、海水晶15g/L，MgCl2·6H2O6.4g/L、CaCl2·2H2O1g/L、MgSO4·7H2O5g/L、KCl2g/L。在接种后24小时时，分别一次性加入外源调控因子或者其组合。利用生物传感仪测定培养基中的葡萄糖浓度，当培养基中的葡萄糖浓度为0g/L时，停止发酵。Schizochytrium B4D1 was inoculated into the fermentation medium, and the components of the fermentation medium were glucose 80g/L, yeast powder 4g/L, soybean peptone 4g/L, sea crystal 15g/L, MgCl 2 6H 2 O6. 4g/L, CaCl 2 2H 2 O 1g/L, MgSO 4 7H 2 O 5g/L, KCl 2g/L. At 24 hours after inoculation, exogenous regulatory factors or a combination thereof were added at one time. Utilize biosensor to measure the glucose concentration in the culture medium, when the glucose concentration in the culture medium is 0g/L, stop fermentation.
实验结果如表3所示：The experimental results are shown in Table 3:
表3table 3
其中细胞干重是指每升发酵液获得的菌体量，角鲨烯含量是指角鲨烯与菌体干重的比例。Wherein, the dry cell weight refers to the amount of cells obtained per liter of fermentation broth, and the squalene content refers to the ratio of squalene to the dry weight of cells.
由表1可以看出，单独加入外源调控因子丁醇或者加入外源调控因子的组合均能增加裂殖壶菌的角鲨烯含量。其中角鲨烯含量以单独加入丁醇8g/L的添加方式为最高。如附图1中，峰1为角鲨烯标准品的检测信号，峰2为添加丁醇后裂殖壶菌油脂样品在相同时间的检测信号。It can be seen from Table 1 that adding the exogenous regulatory factor butanol alone or adding a combination of exogenous regulatory factors can increase the squalene content of Schizochytrium. Wherein the content of squalene is the highest with the addition mode of adding butanol 8g/L alone. As shown in Figure 1, peak 1 is the detection signal of the squalene standard, and peak 2 is the detection signal of the Schizochytrium oil sample at the same time after adding butanol.
这里说明的模块数量和处理规模是用来简化本发明的说明的。对本发明的外源调控因子和菌种及发酵培养基的应用、修改和变化对本领域的技术人员来说是显而易见的。The number of modules and processing scales described here are used to simplify the description of the present invention. The application, modification and variation of the exogenous regulatory factors and strains and fermentation media of the present invention will be obvious to those skilled in the art.
如上所述，根据本发明，由于向发酵培养基中添加了外源调控因子-醇，因此提高了裂殖壶菌中角鲨烯含量，效果明显，可应用于大规模生产。且该方法简单方便，具有经济效益。As mentioned above, according to the present invention, since the exogenous regulatory factor-alcohol is added to the fermentation medium, the content of squalene in Schizochytrium is increased, the effect is obvious, and it can be applied to large-scale production. Moreover, the method is simple and convenient, and has economic benefits.
尽管本发明的实施方案已公开如上，但其并不仅仅限于说明书和实施方式中所列运用，它完全可以被适用于各种适合本发明的领域，对于熟悉本领域的人员而言，可容易地实现另外的修改，因此在不背离权利要求及等同范围所限定的一般概念下，本发明并不限于特定的细节和这里示出与描述的图例。Although the embodiment of the present invention has been disclosed as above, it is not limited to the use listed in the specification and implementation, it can be applied to various fields suitable for the present invention, and it can be easily understood by those skilled in the art Therefore, the invention is not limited to the specific details and examples shown and described herein without departing from the general concept defined by the claims and their equivalents.
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