CN101914581B - 一种提高多烯不饱和脂肪酸发酵产量的方法 - Google Patents
一种提高多烯不饱和脂肪酸发酵产量的方法 Download PDFInfo
- Publication number
- CN101914581B CN101914581B CN201010237946.1A CN201010237946A CN101914581B CN 101914581 B CN101914581 B CN 101914581B CN 201010237946 A CN201010237946 A CN 201010237946A CN 101914581 B CN101914581 B CN 101914581B
- Authority
- CN
- China
- Prior art keywords
- fatty acid
- fermentation
- trehalose
- fermention medium
- unsaturated fatty
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000000855 fermentation Methods 0.000 title claims abstract description 28
- 230000004151 fermentation Effects 0.000 title claims abstract description 28
- 238000000034 method Methods 0.000 title claims abstract description 26
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 title abstract 5
- KWIUHFFTVRNATP-UHFFFAOYSA-N glycine betaine Chemical compound C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 claims abstract description 17
- 241000003595 Aurantiochytrium limacinum Species 0.000 claims abstract description 16
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims abstract description 13
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims abstract description 13
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims abstract description 13
- 229960003237 betaine Drugs 0.000 claims abstract description 13
- 235000021122 unsaturated fatty acids Nutrition 0.000 claims description 14
- 150000004670 unsaturated fatty acids Chemical class 0.000 claims description 10
- -1 polyene unsaturated fatty acid Chemical class 0.000 claims description 6
- 239000000463 material Substances 0.000 abstract description 6
- 244000005700 microbiome Species 0.000 abstract description 6
- 239000001963 growth medium Substances 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 230000008901 benefit Effects 0.000 abstract description 3
- 241000233671 Schizochytrium Species 0.000 abstract 1
- MBMBGCFOFBJSGT-KUBAVDMBSA-N docosahexaenoic acid Natural products CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCC(O)=O MBMBGCFOFBJSGT-KUBAVDMBSA-N 0.000 description 22
- DVSZKTAMJJTWFG-UHFFFAOYSA-N docosa-2,4,6,8,10,12-hexaenoic acid Chemical compound CCCCCCCCCC=CC=CC=CC=CC=CC=CC(O)=O DVSZKTAMJJTWFG-UHFFFAOYSA-N 0.000 description 19
- 210000004027 cell Anatomy 0.000 description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 239000002609 medium Substances 0.000 description 8
- 235000014113 dietary fatty acids Nutrition 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 7
- 229930195729 fatty acid Natural products 0.000 description 7
- 239000000194 fatty acid Substances 0.000 description 7
- 150000004665 fatty acids Chemical class 0.000 description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 229940041514 candida albicans extract Drugs 0.000 description 4
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 4
- 239000002054 inoculum Substances 0.000 description 4
- 235000013923 monosodium glutamate Nutrition 0.000 description 4
- 238000011218 seed culture Methods 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 229940073490 sodium glutamate Drugs 0.000 description 4
- 239000012138 yeast extract Substances 0.000 description 4
- 238000000605 extraction Methods 0.000 description 3
- 230000003204 osmotic effect Effects 0.000 description 3
- 241000199912 Crypthecodinium cohnii Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000607598 Vibrio Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 238000002703 mutagenesis Methods 0.000 description 2
- 231100000350 mutagenesis Toxicity 0.000 description 2
- 230000035790 physiological processes and functions Effects 0.000 description 2
- 238000007670 refining Methods 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000012807 shake-flask culturing Methods 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- YYGNTYWPHWGJRM-UHFFFAOYSA-N (6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyltetracosa-2,6,10,14,18,22-hexaene Chemical compound CC(C)=CCCC(C)=CCCC(C)=CCCC=C(C)CCC=C(C)CCC=C(C)C YYGNTYWPHWGJRM-UHFFFAOYSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 241000233652 Chytridiomycota Species 0.000 description 1
- 241000199914 Dinophyceae Species 0.000 description 1
- WQXNXVUDBPYKBA-UHFFFAOYSA-N Ectoine Natural products CC1=NCCC(C(O)=O)N1 WQXNXVUDBPYKBA-UHFFFAOYSA-N 0.000 description 1
- 208000004232 Enteritis Diseases 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 241000144833 Halomonas salina Species 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 201000004681 Psoriasis Diseases 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 206010047400 Vibrio infections Diseases 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940090949 docosahexaenoic acid Drugs 0.000 description 1
- 235000020669 docosahexaenoic acid Nutrition 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- WQXNXVUDBPYKBA-YFKPBYRVSA-N ectoine Chemical compound CC1=[NH+][C@H](C([O-])=O)CCN1 WQXNXVUDBPYKBA-YFKPBYRVSA-N 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 230000004438 eyesight Effects 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 230000004992 fission Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 230000005923 long-lasting effect Effects 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000008054 signal transmission Effects 0.000 description 1
- TUHBEKDERLKLEC-UHFFFAOYSA-N squalene Natural products CC(=CCCC(=CCCC(=CCCC=C(/C)CCC=C(/C)CC=C(C)C)C)C)C TUHBEKDERLKLEC-UHFFFAOYSA-N 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 1
- 231100000397 ulcer Toxicity 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
- C12P7/6434—Docosahexenoic acids [DHA]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
- C12P7/6432—Eicosapentaenoic acids [EPA]
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明公开了一种提高多烯不饱和脂肪酸发酵产量的方法，以裂殖壶菌为生产菌株发酵生产多烯不饱和脂肪酸，在发酵培养基中添加甘氨酸甜菜碱或海藻糖。本发明，在裂殖壶菌发酵体系中，使用外源甘氨酸甜菜碱或海藻糖处理后，大大提高了裂殖壶菌发酵产生PUFA的产量。本发明通过简单有效的发酵调控，显著提高了微生物产PUFA含量，不危害环境，不增加人力物力，并降低了成本，简单方便且具有经济效益。
Description
技术领域
本发明涉及一种提高多烯不饱和脂肪酸发酵产量的方法，属于生物技术领域。
背景技术
多烯不饱和脂肪酸(PUFA)是细胞和有机体生物膜的重要组成成份，可调节细胞构型、动态平衡、相转变及细胞膜的渗透性，同时还调节与膜有关的生理过程，因此它们可以影响细胞的化学组成、信号传递、免疫及冷适应性，以及与此相关的疾病的发生，PUFA可以转化成调节人体某些生理功能的代谢产物。其中，二十二碳六烯酸(DHA)是细胞膜中PUFA的主要成份，具有重要的生理功能，如增强记忆，提高智力，降低血脂，调节免疫系统等功效，还可以预防和治疗心血管疾病、癌症等。二十二碳五烯酸(DPA)是人类初乳中才有的长链不饱和脂肪酸，是人脑组织、神经细胞的主要组成成份，对婴幼儿神经系统和视力的发育、大脑的形成、记忆力的增强是必不可少的；此外，DPA还可以促进并提高人体的免疫能力。DPA与DHA起协同作用，对II型糖尿病，类风湿性关节炎，牛皮癣，哮喘病，溃疡性大小肠炎等有较大治疗作用，因此具有巨大的商业应用价值。今年来，科学工作者开展了利用海洋微生物发酵生产DHA的研究。常见的微生物包括隐甲藻属、破囊破囊弧菌属等。
相容性溶质是细胞代谢中间产物，无毒，可调节渗透压，防止细胞中离子浓度的激变。当细胞内渗透压激变时，如外部渗透压升高时，细胞开始产生或吸收几种小分子溶质，如海藻糖、甜菜碱、某些氨基酸等以提高细胞内水活度，维持细胞内外渗透压平衡，同时防止细胞水分的流出和盐分的入侵，这些小分子溶质即为“相容性溶质”。
科学工作者开展了利用海洋微生物发酵生产DHA的研究。概括来说，国内已公开的有关专利主要包括以下4个方面：1，关于DHA生产菌株的诱变筛选方法，如中国海洋大学的《海洋真菌裂殖壶菌OUC88的工业应用》(200410075426.X)、南京工业大学的《一种二十二碳六烯酸生产菌株及其诱变筛选方法和其应用》(200910033493.8)等；2，关于培养基的组成，如南京工业大学的《一种裂殖弧菌及利用其生产DHA油脂的方法》(CN200910033869.5)等；3，关于油脂的提取精制，如南京工业大学的《一种从隐甲藻中提取并精制富含DHA脂肪酸的工艺》(200710025079.3)、内蒙古金达威药业有限公司等的《从双鞭甲藻发酵液中提取DHA不饱和脂肪酸的方法》(CN200910159368.1)等；4，关于DHA的应用，如朱阎宏等的《孕产妇营养食品》(CN200610000658.8)、陈义的《即食鱼丸片制备方法》(CN200510045178.9)等。目前，通过简单的发酵调控来增加脂肪酸含量的方法，未发现报道。
国内已公开的有关相容性溶质方面的专利一般包括以下2个方面：1，关于相容性溶质的提取及制备，如申光荣的《一种有机绿色饲料添加剂配制及使用方法》(CN200910109142.0)、阿克佐诺贝尔公司的《甜菜碱的制备方法》(CN00811384.X)、国家海洋局第三海洋研究所的《一种从中性嗜盐菌Halomonas salina检测和提取相容性溶质ectoine的新方法》(CN200610135272.8)等；2，关于相容性溶质的应用。较多的是应用于提高动物的生长性能、植物产量等，如天津生机集团股份有限公司的《一种促进水产动物生长提高肉质的复合制剂及其制备方法》(CN200910307231.6)、卡尔特有限公司的《提高作物产量的方法》(95197919.1、95197917.5等)等；仅有一篇是用于微生物发酵方面：天津科技大学的《一种提高L-谷氨酸发酵产率的新工艺》(CN200910067618.9)。
国外已公开的有关相容性溶质的专利包括：添加相容性溶质提高多肽量，BrianD.Follstad等的《cell culture performance with betaine》(10/226，931)；添加甜菜碱提高乳酸发酵，Shengde Zhou等的《Materials and methods for efficient lactic acid production》(200610109332)。
发明内容
本发明所要解决的技术问题是提供一种简单高效的提高多烯不饱和脂肪酸发酵产量的方法，不危害环境，不增加人力物力，且降低成本。
为解决上述技术问题，本发明采用的技术方案如下：
一种提高多烯不饱和脂肪酸发酵产量的方法，以裂殖壶菌为生产菌株发酵生产多烯不饱和脂肪酸，在发酵培养基中添加相容性溶质。
其中，所述的相容性溶质为甘氨酸甜菜碱或海藻糖。
发酵培养基中，甘氨酸甜菜碱的浓度为10～100mmol/L，优选浓度为10～70mmol/L，最优选浓度为40mmol/L。
发酵培养基中，海藻糖的浓度为10～200mmol/L，优选浓度为40～200mmol/L，优选浓度为80mmol/L。
在发酵培养基中添加少量的外源相容性溶质可以提高海洋微生物发酵生产PUFA的产量。甘氨酸甜菜碱是裂殖壶菌应对环境压力时产生的主要相容性溶质，且比较便宜，约40rmb/kg。海藻糖是微生物应对环境压力时产生的一种相容性溶质，约70rmb/kg。
在发酵条件下，在裂殖壶菌发酵产DHA的培养基中添加甘氨酸甜菜碱或海藻糖，以提高PUFA产量。培养的海洋微生物在遭受不利的发酵条件时，外源相容性溶质是有效用的。这样的外部压力因素包括例如高温、高压、高盐、高渗、低渗、干燥等。经外源相容性溶质处理的裂殖壶菌，能更好地应对外界不断变化的环境，提高DHA的产量。相容性溶质是存留在微生物细胞中的稳定物质，故此，相容性溶质的有益作用是长效的。
有益效果：本发明，在裂殖壶菌发酵体系中，使用外源甘氨酸甜菜碱处理后，大大提高了裂殖壶菌发酵产生PUFA的产量，DPA占总脂肪酸的质量百分比从11.9％提高到16.2％；DHA占总脂肪酸的质量百分比从44.1％提高到49.8％；角鲨烯的质量百分比从0.8％提高到1.7％；而且饱和脂肪酸C14:0和C16:0占总脂肪酸的质量百分比也大幅度降低，分别从10.0％降到5.1％、24.4％降到20％。DHA产量从3.9g/L提高到5.0g/L，提高了28％；DHA占生物量的比(mg/g)从57提高到72；总脂肪酸产量从8.8g/L提高到10g/L。
使用适量海藻糖后，DHA产量从3.9g/L提高到7.5g/L，提高了92％；生物量从60g/L提高到76g/L；DHA占生物量的比(mg/g)从57提高到99；总脂肪酸产量从8.8g/L提高到16.7g/L。
本发明通过简单有效的发酵调控，显著提高了微生物产PUFA含量，不危害环境，不增加人力物力，并降低了成本，简单方便且具有经济效益。
具体实施方式
根据下述实施例，可以更好地理解本发明。然而，本领域的技术人员容易理解，实施例所描述的具体的物料配比、工艺条件及其结果仅用于说明本发明，而不应当也不会限制权利要求书中所详细描述的本发明。
以下实施例的检测方法同《一种裂殖壶菌及利用其生产DHA油脂的方法》(申请号200910033869.5)
实施例1：
菌株为裂殖壶菌HX-308，其保藏编号为CCTCC No.M209059。
种子培养基为：D-葡萄糖40g/L、酵母膏2g/L、谷氨酸钠10g/L、MgCl2 3g/L、CaCl2·2H2O 1g/L、KH2PO4 4g/L、KCl 2g/L、NaCl 15g/L、MgSO4·7H2O 5g/L、FeCl3 0.1g/L。(参考《一种裂殖壶菌及利用其生产DHA油脂的方法》(申请号200910033869.5))。
发酵培养基为：D-葡萄糖40g/L、酵母膏2g/L、谷氨酸钠10g/L、MgCl2 3g/L、(NH4)2SO4 6g/L、KH2PO4 4g/L、KCl 2g/L、NaCl 15g/L、MgSO4·7H2O 5g/L、FeCl3 0.1g/L。(参考《一种裂殖壶菌及利用其生产DHA油脂的方法》(申请号200910033869.5))。
培养方法：将菌种接入种子培养基中，接种量为5％(v/v)；在25℃、170r条件下500mL摇瓶培养24h至对数期，按9％(v/v)的接种量接入添加了一定量甘氨酸甜菜碱(如10mmol/L、40mmol/L、70mmol/L、100mmol/L、200mmol/L)的发酵培养基中培养；发酵至葡萄糖量为0g/L时停止。结果见表1。
表1
实施例2：
菌株为裂殖壶菌HX-308，其保藏编号为CCTCC No.M209059。
种子培养基为：D-葡萄糖40g/L、酵母膏2g/L、谷氨酸钠10g/L、MgCl2 3g/L、CaCl2·2H2O 1g/L、KH2PO4 4g/L、KCl 2g/L、NaCl 15g/L、MgSO4·7H2O 5g/L、FeCl3 0.1g/L。
发酵培养基为：D-葡萄糖40g/L、酵母膏2g/L、谷氨酸钠10g/L、MgCl2 3g/L、(NH4)2SO4 6g/L、KH2PO4 4g/L、KCl 2g/L、NaCl 15g/L、MgSO4·7H2O 5g/L、FeCl3 0.1g/L。
培养方法：将菌种接入种子培养基中，接种量为5％(v/v)；在25℃、170r条件下500mL摇瓶培养24h至对数期，按9％(v/v)的接种量接入添加一定量海藻糖(如10mmol/L、40mmol/L、80mmol/L、160mmol/L、200mmol/L、280mmol/L)的发酵培养基中培养；发酵至葡萄糖量为0g/L时停止。结果见表2。
表2
Claims (5)
1.一种提高多烯不饱和脂肪酸发酵产量的方法，以裂殖壶菌为生产菌株发酵生产多烯不饱和脂肪酸，其特征在于在发酵培养基中添加相容性溶质；
其中，所述的裂殖壶菌为裂殖壶菌HX-308，其保藏编号为CCTCC No.M209059；
所述的相容性溶质为甘氨酸甜菜碱或海藻糖；
发酵培养基中，甘氨酸甜菜碱的浓度为10～100mmol/L；
发酵培养基中，海藻糖的浓度为10～200mmol/L。
2.根据权利要求1所述的提高多烯不饱和脂肪酸发酵产量的方法，其特征在于发酵培养基中，甘氨酸甜菜碱的浓度为10～70mmol/L。
3.根据权利要求2所述的提高多烯不饱和脂肪酸发酵产量的方法，其特征在于发酵培养基中，甘氨酸甜菜碱的浓度为40mmol/L。
4.根据权利要求1所述的提高多烯不饱和脂肪酸发酵产量的方法，其特征在于发酵培养基中，海藻糖的浓度为40～200mmol/L。
5.根据权利要求4所述的提高多烯不饱和脂肪酸发酵产量的方法，其特征在于发酵培养基中，海藻糖的浓度为80mmol/L。
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010237946.1A CN101914581B (zh) | 2010-07-27 | 2010-07-27 | 一种提高多烯不饱和脂肪酸发酵产量的方法 |
| US13/812,496 US20130217085A1 (en) | 2010-07-27 | 2011-01-13 | Methods for improving fermentation yield of polyunsaturated fatty acids |
| PCT/CN2011/070240 WO2012013025A1 (zh) | 2010-07-27 | 2011-01-13 | 提高多烯不饱和脂肪酸发酵产量的方法 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010237946.1A CN101914581B (zh) | 2010-07-27 | 2010-07-27 | 一种提高多烯不饱和脂肪酸发酵产量的方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101914581A CN101914581A (zh) | 2010-12-15 |
| CN101914581B true CN101914581B (zh) | 2012-05-23 |
Family
ID=43322219
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201010237946.1A Active CN101914581B (zh) | 2010-07-27 | 2010-07-27 | 一种提高多烯不饱和脂肪酸发酵产量的方法 |
Country Status (3)
| Country | Link |
|---|---|
| US (1) | US20130217085A1 (zh) |
| CN (1) | CN101914581B (zh) |
| WO (1) | WO2012013025A1 (zh) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101914581B (zh) * | 2010-07-27 | 2012-05-23 | 南京工业大学 | 一种提高多烯不饱和脂肪酸发酵产量的方法 |
| CN102839129A (zh) * | 2011-06-23 | 2012-12-26 | 法国罗凯特兄弟公司 | 一种裂殖壶菌诱变方法及其产生的变异株 |
| CN103305574B (zh) * | 2013-07-16 | 2016-01-27 | 深圳康泰生物制品股份有限公司 | 表达HBsAg的重组酿酒酵母菌发酵培养基及其配制方法和发酵工艺 |
| CN104357498A (zh) * | 2014-09-24 | 2015-02-18 | 江苏省农业科学院 | 一种2,4-二氯苯氧乙酸在提高裂壶藻dha产量及促进裂壶藻油脂积累中的应用 |
| CN105132485B (zh) * | 2015-09-24 | 2019-06-18 | 山东祥维斯生物科技股份有限公司 | 一种裂殖壶菌发酵生产dha的方法 |
| FR3045069B1 (fr) * | 2015-12-14 | 2019-01-25 | Metabolium | Procede d'enrichissement de protistes en lipides riches en acides gras polyinsatures, plus particulierement de classe omega 3, et sa mise en oeuvre pour la production de ces lipides |
| CN114009625B (zh) * | 2021-10-18 | 2024-06-25 | 南京师范大学 | 水产养殖饲料及其制备方法 |
| CN116479063B (zh) * | 2023-05-04 | 2024-02-20 | 厦门汇盛生物有限公司 | 一种ω-3多不饱和脂肪酸的生产方法 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1916156A (zh) * | 2006-07-10 | 2007-02-21 | 温州大学 | 裂殖壶菌(Schizochytrium)WZU4771及其在制备DHA粉末和油脂中的应用 |
| CN101575584A (zh) * | 2009-06-18 | 2009-11-11 | 南京工业大学 | 一种裂殖弧菌及利用其生产dha油脂的方法 |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1549753B1 (en) * | 2002-10-11 | 2010-09-08 | Nippon Suisan Kaisha, Ltd. | Process for producing microbial fat or oil having lowered unsaponifiable matter content |
| WO2007120198A2 (en) * | 2005-11-08 | 2007-10-25 | University Of Florida Research Foundation, Inc. | Materials and methods for improved microbial production of organic compounds |
| WO2007074479A1 (en) * | 2005-12-29 | 2007-07-05 | Abl Biotechnologies Ltd | Novel strain of schizochytrium limacinum useful in the production of lipids and extracellular polysaccharides and process thereof |
| EP2105506A1 (de) * | 2008-03-26 | 2009-09-30 | Lonza Ag | Verfahren zur Herstellung PUFAs enthaltender Öle unter Verwendung von Mikroorganismen der Ordnung Labyrinthulomycota |
| CN101914581B (zh) * | 2010-07-27 | 2012-05-23 | 南京工业大学 | 一种提高多烯不饱和脂肪酸发酵产量的方法 |
-
2010
- 2010-07-27 CN CN201010237946.1A patent/CN101914581B/zh active Active
-
2011
- 2011-01-13 US US13/812,496 patent/US20130217085A1/en not_active Abandoned
- 2011-01-13 WO PCT/CN2011/070240 patent/WO2012013025A1/zh active Application Filing
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1916156A (zh) * | 2006-07-10 | 2007-02-21 | 温州大学 | 裂殖壶菌(Schizochytrium)WZU4771及其在制备DHA粉末和油脂中的应用 |
| CN101575584A (zh) * | 2009-06-18 | 2009-11-11 | 南京工业大学 | 一种裂殖弧菌及利用其生产dha油脂的方法 |
Non-Patent Citations (1)
| Title |
|---|
| 陈诚.裂殖壶菌（Schizochytrium limacinum SR21）发酵制备二十二碳六烯酸（DHA）过程的初步研究.《中国优秀硕士学位论文》.2007,全文. * |
Also Published As
| Publication number | Publication date |
|---|---|
| US20130217085A1 (en) | 2013-08-22 |
| CN101914581A (zh) | 2010-12-15 |
| WO2012013025A1 (zh) | 2012-02-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101914581B (zh) | 一种提高多烯不饱和脂肪酸发酵产量的方法 | |
| Unagul et al. | Coconut water as a medium additive for the production of docosahexaenoic acid (C22: 6 n3) by Schizochytrium mangrovei Sk-02 | |
| DE69621663T2 (de) | Verfahren zur Herstellung von Arachidonsäure | |
| CN108004149B (zh) | 一种海洋原生生物及利用其发酵生产高附加值脂质产品的方法 | |
| US20150104557A1 (en) | Method for the cultivation of microorganisms of the genus thraustochytriales by using an optimized low salt medium | |
| CN102115350B (zh) | 一种桦褐孔菌深层发酵培养基及桦褐孔菌的深层发酵方法 | |
| EP2883958B1 (en) | Improved methods for fermentative production of docosahexaenoic acid | |
| WO2017166831A1 (zh) | 一种ω-7脂肪酸合成物及培养黄丝藻生产该合成物的方法与应用 | |
| CN102864111B (zh) | 一株产二十二碳六烯酸的裂殖壶菌菌株 | |
| CN111117895A (zh) | 藻类脂质组合物和制备及利用所述组合物的方法 | |
| KR102534491B1 (ko) | Dha 및, arg 및 glu 아미노산을 갖는, 트라우스토키트리움 속 미세조류 바이오매스의 농축 방법 | |
| CN102911868B (zh) | 一种微生物培养基及培养方法 | |
| CN103068965A (zh) | 在蔗糖上生长的重组破囊壶菌和其组合物、制备方法及用途 | |
| JP2017077188A (ja) | スクアレンを蓄積した培養微細藻類を含有する魚介類養殖用飼料 | |
| CN110800871A (zh) | 裂壶藻粉在提高反刍动物乳汁中dha含量的应用 | |
| CN103981009A (zh) | 一种裂殖壶菌发酵液破壁提取其胞内油脂的方法 | |
| JP2017502680A (ja) | 微細藻類バイオマスのタンパク質を富化するための方法 | |
| KR20170068461A (ko) | 반연속적 배양 방법 | |
| US20100099901A1 (en) | Method for increasing the content of docosahexaenoic acid in fat-containing materials or in fats and oils | |
| ES2387305T3 (es) | Proceso para el cultivo de microorganismos de la especie Thraustochytrieales | |
| CN107849514A (zh) | 增强木糖的微藻代谢 | |
| CN105349588A (zh) | 利用裂殖壶菌生产二十二碳六烯酸的方法 | |
| CN109275801A (zh) | 添加海洋红酵母提取物的蟹类饲料 | |
| CN112746022A (zh) | 一种预防蝙蝠蛾拟青霉发酵菌丝体中腺苷流失的方法 | |
| CA2655755C (en) | Zygomycetes for fish feed |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230112 Address after: Room 207 and 208, building D6, Jiangsu Life Science Park, No. 9, Weidi Road, Xianlin street, Qixia District, Nanjing, Jiangsu 210000 Patentee after: Nanjing Zhihe Biotechnology Co.,Ltd. Address before: 210009, No. 5, new exemplary Road, Nanjing, Jiangsu Patentee before: Nanjing Tech University |
|
| TR01 | Transfer of patent right | ||
| CP03 | Change of name, title or address |
Address after: Room 207 and 208, building D6, Jiangsu Life Science Park, No. 9, Weidi Road, Xianlin street, Qixia District, Nanjing, Jiangsu 210000 Patentee after: Zhihe Biotechnology (Changzhou) Co.,Ltd. Country or region after: China Address before: Room 207 and 208, building D6, Jiangsu Life Science Park, No. 9, Weidi Road, Xianlin street, Qixia District, Nanjing, Jiangsu 210000 Patentee before: Nanjing Zhihe Biotechnology Co.,Ltd. Country or region before: China |
|
| CP03 | Change of name, title or address |