CN109371071B - Dha微生物油脂中脂肪酸组合物成分调整的方法 - Google Patents
Dha微生物油脂中脂肪酸组合物成分调整的方法 Download PDFInfo
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Abstract
本发明涉及微生物发酵技术领域,公开了一种DHA微生物油脂中脂肪酸组合物成分调整的方法,在发酵的第0~80小时,将培养基的碳氮比控制在3~20:1,将发酵温度控制在29~32℃;在发酵的第81~120小时,将培养基的碳氮比控制在1~15:1将发酵温度控制在19~28℃的温度条件。随着氮源的消耗、再控制碳源的补充,营造一个营养缺陷型生长环境,有助于脂肪酸的转化。精炼阶段采用变温与程序控温的冬化工艺,去除相对饱和度高的脂肪酸含量,提高多不饱和脂肪酸的相对含量,改善油脂的低温流动性能。该方法取得的组合物可用于制备婴幼儿配方食品、保健品、健康食品以及普通食品。
Description
技术领域
本发明涉及微生物发酵技术领域,具体涉及一种DHA微生物油脂中脂肪酸组合物成分调整的方法。
背景技术
微生物油脂,又称为单细胞油脂,是由酵母、霉菌和微藻等产脂微生物在一定条件下利用碳源、氮源、微量元素,在菌体内产生的大量油脂。
产油微生物资源丰富,能在多种培养条件下生长,进行工业规模生产和开发有着巨大的潜力。微生物油脂的多不饱和脂肪酸含量较高,其中的二十二碳六烯酸(DHA)、花生四烯酸(ARA)等为人体必须脂肪酸,具有重要的生理功能。
婴儿缺乏ARA和DHA,可造成永久性智力低下和视力障碍,6~12岁儿童的皮肤瘙痒、眼角干燥、上课注意力不集中;对高血压、高血脂、糖尿病、病毒感染等疾病的预防和治疗也有显著的效果。
市场销售的DHA、ARA等产品,主要从深海鱼油中提取,由于原料的限制,无法进行规模化生产;同时不饱和脂肪酸含量不稳定,而且产量低、成本高。因此,微生物油脂已成为获取高附加值脂肪酸,如亚麻酸(GLA)、花生四烯酸(ARA)、二十碳五烯酸(EPA)、二十二碳六烯酸(DHA)等的重要原料。
但现有的研究表明,过量的EPA对胎儿和婴幼儿的生长、代谢存在不良的影响,因此对孕妇和婴幼儿食用的微生物油脂,应严格控制其中的EPA含量。
由于吾肯氏菌属、裂殖壶菌属、破囊壶菌目、隐甲藻属、酵母等微生物发酵生产的微生物油脂DHA含量较高,是生产DHA油脂所主要使用的菌种。而现有的利用吾肯氏菌属、裂殖壶菌属、破囊壶菌目、隐甲藻属、酵母等微生物发酵生产微生物油脂的方法所获取的油脂中,DHA的含量还不够高,油脂中还含有较多的EPA和一些有害脂肪酸,如豆蔻酸、月桂酸、芥酸等,影响了微生物油脂的使用。
发明内容
本发明的目的是为了克服现有技术生产的DHA微生物油脂中有害脂肪酸含量偏高、以及DHA的含量不够高、EPA含量较高的问题,提供一种DHA微生物油脂中脂肪酸组合物成份调整的方法,该方法利用吾肯氏菌属、裂殖壶菌属、破囊壶菌目、隐甲藻属、酵母等微生物发酵生产DHA微生物油脂,可以调控微生物油脂中脂肪酸组合物的含量,生产高DHA含量的微生物油脂,且其中的有害脂肪酸含量很低。
为了实现上述目的,第一方面,本发明提供了一种DHA微生物油脂中脂肪酸组合物成分调整的方法,包括在菌种发酵的第0~80小时,将培养基的碳氮比控制在3~20:1,培养温度控制在29~32℃之间;在菌种发酵的第81小时以后,将培养基的碳氮比控制在1~15:1,培养温度控制在19~28℃之间。
优选地,所述控制碳氮比的方式为,控制培养基中葡萄糖和谷氨酸钠的含量,以维持所需的碳氮比。
优选地,所述控制培养基中葡萄糖和谷氨酸钠含量的过程为,在发酵的第0~16小时,葡萄糖含量为2~8g/100ml,谷氨酸钠的含量为1~5g/100ml;第17~55小时,葡萄糖含量为2~6g/100ml,谷氨酸钠的含量为1~3g/100ml;第56~80小时,葡萄糖含量为1~5g/100ml,谷氨酸钠的含量为1~2g/100ml;第81~95小时,葡萄糖含量为0.5~1.0g/100ml,谷氨酸钠的含量为0.75~1.0g/100ml;第95小时以后,葡萄糖含量为<0.5g/100ml,谷氨酸钠的含量为0.5~0.75g/100ml。
优选地,本发明提供的DHA微生物油脂中脂肪酸组合物成分调整的方法,还包括在微生物油脂的精制过程中,先将油脂升温至70~90℃脱水,然后逐渐冷却至20~30℃常温冬化16~24小时,过滤、脱水后,再按设定的程序逐渐降温到-10~1℃保温,低温冬化48~90小时。
第二方面,本发明提供了一种DHA微生物油脂,根据本发明第一方面所述的任一种DHA微生物油脂中脂肪酸组合物成分调整的方法制得,以微生物油脂的总量为基准,含有以下组分:不低于40wt%的DHA,不超过3wt%的EPA;0~5wt%的C12:0脂肪酸;0~8wt%的C14:0、C14:1脂肪酸;5~50wt%的C16:0和C16:1脂肪酸;0.5~10wt%的C18:0、C18:1和C18:2脂肪酸;0~5wt%的C22:1n9脂肪酸。
优选地,本发明的DHA微生物油脂中,DHA与EPA的比例为13.3~600:1。
优选地,本发明的DHA微生物油脂中不饱和脂肪酸与饱和脂肪酸的含量比不低于0.6。
进一步优选地,本发明的DHA微生物油脂中不饱和脂肪酸与饱和脂肪酸的含量比不低于2.3。
第三方面,本发明提供了一种组合物,包含本发明第二方面所述的DHA微生物油脂,所述组合物适用于制作婴幼儿配方食品(婴幼儿配方奶粉)、保健食品、健康食品以及普通食品等。
通过上述技术方案,不仅微生物油脂的产量较高,微生物油脂中DHA的含量高、EPA的含量低,有害脂肪酸豆蔻酸、月桂酸和芥酸的含量很低,可在-10~5℃条件下可保障澄清透明状态。
具体实施方式
在本文中所披露的范围的端点和任何值都不限于该精确的范围或值,这些范围或值应当理解为包含接近这些范围或值的值。对于数值范围来说,各个范围的端点值之间、各个范围的端点值和单独的点值之间,以及单独的点值之间可以彼此组合而得到一个或多个新的数值范围,这些数值范围应被视为在本文中具体公开。
在本发明的实施例和对照例中,DHA微生物油脂的发酵菌种均使用裂壶藻(Schizochytrium),所用菌种和各种培养用品均由市售而得,所涉及的试剂和检测方法均按国家标准执行。
实施例1
1)生产菌种:将原始菌种接入灭菌冷却好的培养基中,在振荡摇瓶中培养。
摇瓶培养基配方为:葡萄糖4.5%,谷氨酸钠3.2%,酵母浸膏0.62%,氯化钠1.8%,磷酸二氢钾0.7%,硫酸镁0.55%,氯化钙0.03%,微量元素(硫酸镍、硫酸铜、钼酸钠、氯化锰、氯化钴、硫酸锌、硫酸亚铁等)0.1%,维生素(B2、B6、B12等)0.1%。
培养条件为:200转/分摇床上,28±1℃培养36~44小时,待菌丝长好后取下,并瓶后接入一级种子罐。
2)一级种子:将菌种接入灭菌冷却好的培养基中,在一级种子罐中培养。
培养基的配方为:葡萄糖3%,酵母粉0.83%,谷氨酸钠0.62%,酵母浸膏0.62%,氯化钠0.62%,磷酸二氢钾0.7%,硫酸镁0.52%,氯化钙0.02%,碳酸氢钠0.02%,硫酸钠0.93%,硫酸氨0.10%,氯化钾0.08%,微量元素(成分同上)0.10%,维生素(成分同上)0.10%,环氧硅醚0.03%。
培养条件为:温度30±2℃,转速90rpm,通气量为0.50vvm,培养18~24小时。
3)二级种子:将4%的一级菌种接入灭菌冷却好的培养基中,在二级种子罐中培养。
培养基的配方为:葡萄糖5%,谷氨酸钠2.0%,酵母浸膏1.0%,氯化钠0.14%,磷酸二氢钾0.16%,硫酸镁0.50%,氯化钙0.02%,碳酸氢钠0.02%,硫酸钠0.91%,硫酸氨0.10%,氯化钾0.08%,微量元素(成分同上)0.09%,维生素(成分同上)0.09%,环氧硅醚0.36%。
培养条件为:温度30±2℃、转速90rpm,培养14~16小时。
4)主发酵:将二级种子液按10%的比例接种于发酵罐的发酵培养基中进行发酵。
培养基配方为:葡萄糖5%,谷氨酸钠2.5%,酵母浸膏1.0%,氯化钠0.29%,磷酸二氢钾0.28%,硫酸镁0.66%,氯化钙0.03%,碳酸氢钠0.02%,硫酸钠0.58%,硫酸氨0.13%,氯化钾0.11%,微量元素(成分同上)0.13%,维生素(成分同上)0.13%,环氧硅醚0.03%。
培养条件为:转速90转/分,0~80小时温度控制在30℃,80小时以后温度控制在22℃;通气量为0.5VVm;用含50%葡萄糖的糖罐添加葡萄糖、添加浓度为60%的谷氨酸钠溶液调整培养基中葡萄糖和谷氨酸钠的含量来控制碳氮比,培养基中葡萄糖和谷氨酸钠的含量及PH的控制见表1,培养5天。
表1
5)提取毛油:发酵完成后,用三相离心机脱水,将菌丝体与水分离、用95%的乙醇脱水后再加入己烷萃取得到毛油。
6)水洗脱胶:加入油重10%的纯净水,加热到85℃,以80rpm的转速搅拌20分钟,静置2小时后分水。
7)酸炼:将油加热到75℃,加入占油重4‰的柠檬酸,以80rpm的转速搅拌40分钟。再加入油重10%的85℃热水,搅拌20分钟,静置3小时后分水。
8)碱炼:将油加热到45℃,根据油脂酸价计算加碱量(加碱量=7.13×10-4×酸价×油重),加入40%的氢氧化钠溶液,以80rpm的转速搅拌50分钟;将油脂温度升温到80℃,再加入油重5%的85℃纯净水,以80rpm的速度搅拌15分钟,用二相离心机离心去除皂角。
9)脱水:碱炼后的油脂加热到80℃,在-0.1MPa负压下脱水35分钟。
10)常温冬化:脱水后的油脂自然降温,油温降至25℃保持养晶。常温冬化时间20小时。
11)过滤:经常温冬化后的油脂,使用板框压滤机过滤,过滤介质为工业滤布,过滤压力在0.3MPa。
12)再脱水:将经过滤后的油脂加热到80℃,在-0.1MPa负压下脱水35分钟。
13)低温冬化:将脱水后的油脂按设定程序降温:先将油脂以每半小时10℃的速度快速降温,达到45℃时以每小时降温3℃,逐渐减少为每小时降温1℃。油温降至14℃时开始回温养晶,回温温度1.5℃,回温时间4.5小时,回温后以每小时1.5℃的速度继续降温至-5℃保温,低温冬化时间70小时。
14)再过滤:经常温冬化后的油脂,使用板框压滤机过滤,过滤介质为工业滤布,过滤压力在0.2MPa。
15)脱色:添加1.5%活性炭、1.5%活性白土,脱色70分钟。
16)脱臭:脱臭温度在175±2℃时开始计时,脱臭温度维持在175±2℃,脱臭时间为4小时,蒸汽压力维持在0.2MPa~0.3MPa,真空度维持在50Pa,蒸汽耗量控制在油重5%左右。降温、破真空,得到成品油脂。
实施例2
1)生产菌种:将原始菌种接入灭菌冷却好的培养基中,在振荡摇瓶中培养。
摇瓶培养基配方为:葡萄糖4.5%,谷氨酸钠3.2%,酵母浸膏0.62%,氯化钠1.8%,磷酸二氢钾0.7%,硫酸镁0.55%,氯化钙0.03%,微量元素(硫酸镍、硫酸铜、钼酸钠、氯化锰、氯化钴、硫酸锌、硫酸亚铁等)0.1%,维生素(B2、B6、B12等)0.1%。
培养条件为:200转/分摇床上,28±1℃培养36~44小时,待菌丝长好后取下,并瓶后接入一级种子罐。
2)一级种子:将菌种接入灭菌冷却好的培养基中,在一级种子罐中培养。
培养基的配方为:葡萄糖3%,酵母粉0.83%,谷氨酸钠0.62%,酵母浸膏0.62%,氯化钠0.62%,磷酸二氢钾0.7%,硫酸镁0.52%,氯化钙0.02%,碳酸氢钠0.02%,硫酸钠0.93%,硫酸氨0.10%,氯化钾0.08%,微量元素(成分同上)0.10%,维生素(成分同上)0.10%,环氧硅醚0.03%。
培养条件为:温度30±2℃,转速90转/分,通气量为0.5vvm,培养18~22小时。
3)二级种子:将4%的一级菌种接入灭菌冷却好的培养基中,在二级种子罐中培养。
培养基的配方为:葡萄糖5%,谷氨酸钠2.0%,酵母浸膏1.0%,氯化钠0.14%,磷酸二氢钾0.16%,硫酸镁0.50%,氯化钙0.02%,碳酸氢钠0.02%,硫酸钠0.91%,硫酸氨0.10%,氯化钾0.08%,微量元素(成分同上)0.09%,维生素(成分同上)0.09%,环氧硅醚0.36%。
培养条件为:温度30±2℃、转速90转/分,培养14~16小时。
4)主发酵:将二级种子液按10%的比例接种于发酵罐的发酵培养基中进行发酵。
培养基配方为:葡萄糖5%,谷氨酸钠2.5%,酵母浸膏1.0%,氯化钠0.29%,磷酸二氢钾0.28%,硫酸镁0.66%,氯化钙0.03%,碳酸氢钠0.02%,硫酸钠0.58%,硫酸氨0.13%,氯化钾0.11%,微量元素(成分同上)0.13%,维生素(成分同上)0.13%,环氧硅醚0.03%。
培养条件为:转速90转/分,0~80小时温度控制为29℃,80小时以后温度控制在19℃;通气量为0.45VVm;用含50%葡萄糖的糖罐添加葡萄糖、添加60%的谷氨酸钠溶液调整培养基中葡萄糖和谷氨酸钠的含量来控制碳氮比,培养基中葡萄糖和谷氨酸钠的含量及PH的控制见表2,培养5天。
表2
5)提取毛油:发酵完成后,用三相离心机脱水,将菌丝体与水分离、用95%的乙醇脱水后再加入己烷萃取得到毛油。
6)水洗脱胶:加入油重10%的纯净水,加热到85℃,以80rpm的转速搅拌20分钟,静置2小时后分水。
7)酸炼:将油加热到75℃,加入占油重4‰的柠檬酸,以80rpm的转速搅拌40分钟。再加入油重10%的85℃热水,搅拌20分钟,静置3小时后分水。
8)碱炼:将油加热到45℃,根据油脂酸价计算加碱量(加碱量=7.13×10-4×酸价×油重),加入40%的氢氧化钠溶液,以80rpm的转速搅拌50分钟;将油脂温度升温到80℃,再加入油重5%的85℃纯净水,以80rpm的速度搅拌15分钟,用二相离心机离心去除皂角。
9)脱水:碱炼后的油脂加热到70℃,在-0.1MPa负压下脱水35分钟。
10)常温冬化:脱水后的油脂自然降温,油温降至20℃保持养晶。常温冬化时间16小时。
11)过滤:经常温冬化后的油脂,使用板框压滤机过滤,过滤介质为工业滤布,过滤压力在0.3MPa。
12)再脱水:将经过滤后的油脂加热到80℃,在-0.1MPa负压下脱水35分钟。
13)低温冬化:将脱水后的油脂按设定程序降温:先将油脂以每半小时10℃的速度快速降温,达到45℃时以每小时降温3℃,逐渐减少为每小时降温1℃。油温降至15℃时开始回温养晶,回温温度1℃,回温时间4小时,回温后以每小时2℃的速度继续降温至1℃保温,低温冬化时间48小时。
14)再过滤:经常温冬化后的油脂,使用板框压滤机过滤,过滤介质为工业滤布,过滤压力在0.2MPa。
15)脱色:添加1.5%活性炭、1.5%活性白土,脱色70分钟。
16)脱臭:脱臭温度在175±2℃时开始计时,脱臭温度维持在175±2℃,脱臭时间为4小时,蒸汽压力维持在0.2MPa~0.3MPa,真空度维持在50Pa,蒸汽耗量控制在油重5%左右。降温、破真空,得到成品油脂。
实施例3
1)生产菌种:将原始菌种接入灭菌冷却好的培养基中,在振荡摇瓶中培养。
摇瓶培养基配方为:葡萄糖4.5%,谷氨酸钠3.2%,酵母浸膏0.62%,氯化钠1.8%,磷酸二氢钾0.7%,硫酸镁0.55%,氯化钙0.03%,微量元素(硫酸镍、硫酸铜、钼酸钠、氯化锰、氯化钴、硫酸锌、硫酸亚铁等)0.1%,维生素(B2、B6、B12等)0.1%。
培养条件为:200转/分摇床上,28±1℃培养36~44小时,待菌丝长好后取下,并瓶后接入一级种子罐。
2)一级种子:将菌种接入灭菌冷却好的培养基中,在一级种子罐中培养。
培养基的配方为:葡萄糖3%,酵母粉0.83%,谷氨酸钠0.62%,酵母浸膏0.62%,氯化钠0.62%,磷酸二氢钾0.7%,硫酸镁0.52%,氯化钙0.02%,碳酸氢钠0.02%,硫酸钠0.93%,硫酸氨0.10%,氯化钾0.08%,微量元素(成分同上)0.10%,维生素(成分同上)0.10%,环氧硅醚0.03%。
培养条件为:温度30±2℃,转速90转/分,通气量为0.50vvm,培养18~22小时。
3)二级种子:将4%的一级菌种接入灭菌冷却好的培养基中,在二级种子罐中培养。
培养基的配方为:葡萄糖5%,谷氨酸钠2.0%,酵母浸膏1.0%,氯化钠0.14%,磷酸二氢钾0.16%,硫酸镁0.50%,氯化钙0.02%,碳酸氢钠0.02%,硫酸钠0.91%,硫酸氨0.10%,氯化钾0.08%,微量元素(成分同上)0.09%,维生素(成分同上)0.09%,环氧硅醚0.36%。
培养条件为:温度30±2℃、转速90转/分,培养14~16小时。
4)主发酵:将二级种子液按10%的比例接种于发酵罐的发酵培养基中进行发酵。
培养基配方为:葡萄糖5%,谷氨酸钠2.5%,酵母浸膏1.0%,氯化钠0.29%,磷酸二氢钾0.28%,硫酸镁0.66%,氯化钙0.03%,碳酸氢钠0.02%,硫酸钠0.58%,硫酸氨0.13%,氯化钾0.11%,微量元素(成分同上)0.13%,维生素(成分同上)0.13%,环氧硅醚0.03%。
培养条件为:转速90转/分,0~80小时温度控制在32℃,80小时以后温度控制在28℃;通气量为0.5VVm;用含50%葡萄糖的糖罐添加葡萄糖、添加60%的谷氨酸钠溶液调整培养基中葡萄糖和谷氨酸钠的含量来控制碳氮比,培养基中葡萄糖和谷氨酸钠的含量及PH的控制见表3,培养5天。
表3
5)提取毛油:发酵完成后,用三相离心机脱水,将菌丝体与水分离、用95%的乙醇脱水后再加入己烷萃取得到毛油。
6)水洗脱胶:加入油重10%的纯净水,加热到85℃,以80rpm的转速搅拌20分钟,静置2小时后分水。
7)酸炼:将油加热到75℃,加入占油重4‰的柠檬酸,以80rpm的转速搅拌40分钟。再加入油重10%的85℃热水,搅拌20分钟,静置3小时后分水。
8)碱炼:将油加热到45℃,根据油脂酸价计算加碱量(加碱量=7.13×10-4×酸价×油重),加入40%的氢氧化钠溶液,以80rpm的转速搅拌50分钟;将油脂温度升温到80℃,再加入油重5%的85℃纯净水,以80rpm的速度搅拌15分钟,用二相离心机离心去除皂角。
9)脱水:将经水洗脱胶、碱炼脱皂后的油脂加热到90℃,在-0.1MPa负压下脱水35分钟。
10)常温冬化:脱水后的油脂自然降温,油温降至30℃保持养晶。常温冬化时间24小时。
11)过滤:经常温冬化后的油脂,使用板框压滤机过滤,过滤介质为工业滤布,过滤压力在0.3MPa。
12)再脱水:将经过滤后的油脂加热到85℃,在-0.1MPa负压下脱水35分钟。
13)低温冬化:将脱水后的油脂按设定程序降温:先将油脂以每半小时10℃的速度快速降温,达到45℃时以每小时降温3℃,逐渐减少为每小时降温1℃。油温降至13℃时开始回温养晶,回温温度2℃,回温时间5小时,回温后以每小时1℃的速度继续降温至-10℃,确保养晶时间不少于16小时。低温冬化时间90小时。
14)再过滤:经常温冬化后的油脂,使用板框压滤机过滤,过滤介质为工业滤布,过滤压力在0.2MPa。
15)脱色:添加1.5%活性炭、1.5%活性白土,脱色70分钟。
16)脱臭:脱臭温度在175±2℃时开始计时,脱臭温度维持在175±2℃,脱臭时间为4小时,蒸汽压力维持在0.2MPa~0.3MPa,真空度维持在50Pa,蒸汽耗量控制在油重5%左右。降温、破真空,得到成品油脂。
对照例
参照实施例1的方法培养菌种和发酵,不同的是主发酵培养基中含葡萄糖4.5%,谷氨酸钠2%,酵母浸膏0.9%,(碳氮比为10.7:1),采用28℃的恒温培养5天;低温冬化时快速降温至0℃结晶养晶48小时。
实验结果
分别将实施例1、实施例2、实施例3和对照例得到的成品油脂,气相色谱检测其中的脂肪酸成分,各脂肪酸在成品油脂中所占比例见表4。
表4
通过表4的结果可以看出,采用本发明的DHA微生物油脂中脂肪酸组合物成分调整的方法实施例DHA含量更高,有害脂肪酸含量更低。另外还具有更好的低温凝固性能。
本发明提供的微生物油脂中DHA含量高、EPA含量低,有害脂肪酸的含量低,低温凝固性能好,可用于制作婴幼儿配方食品,尤其是婴幼儿配方奶粉;还可制作成保健品,根据本领域的技术人员已知的DHA与许多疾病的关系提供给有相关需求的人群作治疗、保健要求用;还可制作成健康食品与普通食品等为人体提供所需的营养,补充日常摄入的不足。
以上详细描述了本发明的优选实施方式,但是,本发明并不限于此。在本发明的技术构思范围内,可以对本发明的技术方案进行多种简单变型,包括各个技术特征以任何其它的合适方式进行组合,这些简单变型和组合同样应当视为本发明所公开的内容,均属于本发明的保护范围。
Claims (1)
1.一种DHA微生物油脂中脂肪酸组合物成分调整的方法,其特征在于,包括在菌种发酵的第0~80小时,将培养基的碳氮比控制在3~20:1,培养温度控制在29~32℃之间;在菌种发酵的第81小时以后,将培养基的碳氮比控制在1~15:1,培养温度控制在19~28℃之间,以提高DHA的转化率,降低其它脂肪酸的含量,所述菌种均使用裂壶藻;
所述控制碳氮比的方式为,控制培养基中葡萄糖和谷氨酸钠的含量,以维持所需的碳氮比,所述控制培养基中葡萄糖和谷氨酸钠含量的过程为,在发酵的第0~16小时,葡萄糖含量为2~8g/100ml,谷氨酸钠的含量为1~5g/100ml;第17~55小时,葡萄糖含量为2~6g/100ml,谷氨酸钠的含量为1~3g/100ml;第56~80小时,葡萄糖含量为1~5g/100ml,谷氨酸钠的含量为1~2g/100ml;第81~95小时,葡萄糖含量0.5~1.0g/100ml,谷氨酸钠的含量为0.75~1.0g/100ml;第96小时以后,葡萄糖含量<0.5g/100ml,谷氨酸钠的含量为0.5~0.75g/100ml;以及
所述DHA微生物油脂中脂肪酸组合物成分调整的方法还包括在微生物油脂的精制过程中,先将油脂升温至70~90℃脱水,然后逐渐冷却至20~30℃常温冬化16~24小时,过滤、脱水后,再按设定的程序逐渐降温到-10~1℃保温,低温冬化48~90小时。
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CN104131044A (zh) * | 2014-06-13 | 2014-11-05 | 顾慧雅 | 一种回收利用微生物提油后的生物质来生产不饱和脂肪酸的方法 |
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WO2016059540A1 (en) * | 2014-10-16 | 2016-04-21 | MARA Renewables Corporation | Semi-continuous culture methods |
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CN105018539A (zh) * | 2015-08-24 | 2015-11-04 | 青岛旭能生物工程有限责任公司 | 一种培养裂殖壶菌高产dha的方法 |
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