高产多不饱和脂肪酸发酵条件优化的研究
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摘要
脂肪酸是人类的重要营养源,而多不饱和脂肪酸(Polyunsaturated fatty acids,PUFAs)对人类正常生命活动所发挥的作用更为重要,它们不仅是维持人类正常生命活动所必需的,而且对人类很多疾病具有明显的预防和治疗作用。微生物发酵法生产多不饱和脂肪酸不受原料、气候、产地的限制,易规模化、产业化生产等优点,因此具有广阔的市场前景。
本课题主要研究如下:(1)从保藏的深黄被孢霉(Mortierella isabellina)菌种中分离出供试验用菌株;(2)发酵生产多不饱和脂肪酸培养基的优化;(3)发酵生产多不饱和脂肪酸条件的优化;(4)深黄被孢霉菌油的理化性质检测。通过对深黄被孢霉产脂培养基的初步筛选后,最终确定3号培养基和8号培养基为优化后的培养基。用气相色谱分析8号培养基和3号培养基的菌油后,发现8号培养基和3号培养基中饱和脂肪酸含量分别为14.50%、14.02%,不饱和脂肪酸含量为85.21%、85.45%,不饱和脂肪酸占总脂肪酸的含量分别为85.46%、85.91%。8号培养基中亚麻酸(C18:3n3)含量为8.07%,其中γ-亚麻酸(C18:3n6,Gamma-Linolenic acid,GLA)的含量为6.80%;3号培养基中亚麻酸含量为8.97%,其中GLA的含量为6.50%。
通过培养基优化试验和条件的初步优化试验后,均显示了8号培养基优于3号培养基,因此只选择了对8号培养基的发酵条件进行进一步的优化实验。将8号培养基在1号优化条件和2号优化条件下发酵后,最终确定的优化条件为1号优化条件。用气相色谱分析检测8号培养基在1号优化条件下的两个菌油样品的脂肪酸组成,可以得到脂肪酸的平均组成分别为:棕榈酸(C16:0)9.65%,硬脂酸(C18:0)2.05%,油酸(C18:1n9)50.00%,亚油酸(C18:2n6)23.35%,α-亚麻酸(C18:3n3)1.35%,GLA 8.65%,花生酸(C20:0)0.40%,花生一烯酸(C20:1)0.69%,花生二烯酸(C20:2)0.35%,二十四碳一烯酸(C24:1(n-6))0.60%,不饱和脂肪酸占总脂肪酸的平均含量为87.03%。
经检测,发现深黄被孢霉菌毛油除酸价偏高外,其它的理化性质与推荐的微生物油脂标准相差不大,这就说明了此菌油经精练处理后可以用来食用。
Fatty acid is important nutrient source.Whereas polyunsaturated fatty acids play a more important role in humanity normal vital movement.They are not only necessary for maintaining vital movement but prevent and cure a great many sickness for human.Producing PUFAs by fermentation is free from raw material、climate、producing area' restrict,and has some merits,for exameple,being easy to produce in a big way and industrialization produce and so on.
Some aspects were studied in this subject included:(1) isolating test strain from conservative Mortierella isabellina,(2) optimizing culture medium for highly productive PUFAs by fermentation,(3) optimizing fermentation conditions for highly productive PUFAs,(4) checking and measuring physical and chemical characters from Mortierella isabellina strain oil.
By primary screening of Mortierella isabellina producing fat culture medium, No 3 culture medium and No 8 culture medium were finally choosed as optimized culture medium.Analyzing the strain oil of No 8 culture medium and No 3 culture medium by gas chromatography,it showed that saturated fatty acids in fatty acids of the Mortierella isabellina oil was 14.50% and unsaturated fatty acids in fatty acids of the Mortierella isabellina oil was 85.21% and unsaturated fatty acids in the total fatty acids was 85.46% in the No 8 culture medium,and saturated fatty acids in fatty acids of the Mortierella isabellina oil was 14.02% and unsaturated fatty acids in fatty acids of the Mortierella isabellina oil was 85.45% and unsaturated fatty acids in the total fatty acids was 85.91% in the No 3 culture medium.No 8 culture medium yielded 8.07% C18:3n3 and 6.80% in the oil contend was C18:3n6; No 3 culture medium yielded 8.97% C18:3n3 and 6.50% in the oil contend was C18:3n6.
It showed that No 8 culture medium was better than No 3 culture medium by the primary optimized culture medium and fermentation conditions,and No 8 culture medium was ultimately selected to be more step optimized conditions try.After No 8 culture medium was fermented in No 1 opotimized condition and No 2 opotimized condition,No 1 opotimized condition was selected as opotimized condition because of the oil yield in No 2 opotimized condition lower it in No 1 opotimized condition.Checking and measuring the fatty acid composition of No 8 culture medium in No 1 opotimized condition by gas chromatography,it showed that average fatty acid composition was C16:0 9.65%,C18:0 2.05%,C18:1n9 50.00%, C18:2n6 23.35%,C18:3n3 1.35%,C18:3n6 8.65%,C20:0 0.40%,C20:1 0.69%,C20:2n6 0.35%,C24:1(n-6) 0.60%,average contend of unsaturated fatty acids in the total fatty acids was 87.03%.
Through checking and measuring Mortierella isabellina gross oil , we discovered that other physical and chemical characters were in accord with recommended the microbial oil standard other than higher acid value.This showed that the refined strain oil was edible.
本课题主要研究如下:(1)从保藏的深黄被孢霉(Mortierella isabellina)菌种中分离出供试验用菌株;(2)发酵生产多不饱和脂肪酸培养基的优化;(3)发酵生产多不饱和脂肪酸条件的优化;(4)深黄被孢霉菌油的理化性质检测。通过对深黄被孢霉产脂培养基的初步筛选后,最终确定3号培养基和8号培养基为优化后的培养基。用气相色谱分析8号培养基和3号培养基的菌油后,发现8号培养基和3号培养基中饱和脂肪酸含量分别为14.50%、14.02%,不饱和脂肪酸含量为85.21%、85.45%,不饱和脂肪酸占总脂肪酸的含量分别为85.46%、85.91%。8号培养基中亚麻酸(C18:3n3)含量为8.07%,其中γ-亚麻酸(C18:3n6,Gamma-Linolenic acid,GLA)的含量为6.80%;3号培养基中亚麻酸含量为8.97%,其中GLA的含量为6.50%。
通过培养基优化试验和条件的初步优化试验后,均显示了8号培养基优于3号培养基,因此只选择了对8号培养基的发酵条件进行进一步的优化实验。将8号培养基在1号优化条件和2号优化条件下发酵后,最终确定的优化条件为1号优化条件。用气相色谱分析检测8号培养基在1号优化条件下的两个菌油样品的脂肪酸组成,可以得到脂肪酸的平均组成分别为:棕榈酸(C16:0)9.65%,硬脂酸(C18:0)2.05%,油酸(C18:1n9)50.00%,亚油酸(C18:2n6)23.35%,α-亚麻酸(C18:3n3)1.35%,GLA 8.65%,花生酸(C20:0)0.40%,花生一烯酸(C20:1)0.69%,花生二烯酸(C20:2)0.35%,二十四碳一烯酸(C24:1(n-6))0.60%,不饱和脂肪酸占总脂肪酸的平均含量为87.03%。
经检测,发现深黄被孢霉菌毛油除酸价偏高外,其它的理化性质与推荐的微生物油脂标准相差不大,这就说明了此菌油经精练处理后可以用来食用。
Fatty acid is important nutrient source.Whereas polyunsaturated fatty acids play a more important role in humanity normal vital movement.They are not only necessary for maintaining vital movement but prevent and cure a great many sickness for human.Producing PUFAs by fermentation is free from raw material、climate、producing area' restrict,and has some merits,for exameple,being easy to produce in a big way and industrialization produce and so on.
Some aspects were studied in this subject included:(1) isolating test strain from conservative Mortierella isabellina,(2) optimizing culture medium for highly productive PUFAs by fermentation,(3) optimizing fermentation conditions for highly productive PUFAs,(4) checking and measuring physical and chemical characters from Mortierella isabellina strain oil.
By primary screening of Mortierella isabellina producing fat culture medium, No 3 culture medium and No 8 culture medium were finally choosed as optimized culture medium.Analyzing the strain oil of No 8 culture medium and No 3 culture medium by gas chromatography,it showed that saturated fatty acids in fatty acids of the Mortierella isabellina oil was 14.50% and unsaturated fatty acids in fatty acids of the Mortierella isabellina oil was 85.21% and unsaturated fatty acids in the total fatty acids was 85.46% in the No 8 culture medium,and saturated fatty acids in fatty acids of the Mortierella isabellina oil was 14.02% and unsaturated fatty acids in fatty acids of the Mortierella isabellina oil was 85.45% and unsaturated fatty acids in the total fatty acids was 85.91% in the No 3 culture medium.No 8 culture medium yielded 8.07% C18:3n3 and 6.80% in the oil contend was C18:3n6; No 3 culture medium yielded 8.97% C18:3n3 and 6.50% in the oil contend was C18:3n6.
It showed that No 8 culture medium was better than No 3 culture medium by the primary optimized culture medium and fermentation conditions,and No 8 culture medium was ultimately selected to be more step optimized conditions try.After No 8 culture medium was fermented in No 1 opotimized condition and No 2 opotimized condition,No 1 opotimized condition was selected as opotimized condition because of the oil yield in No 2 opotimized condition lower it in No 1 opotimized condition.Checking and measuring the fatty acid composition of No 8 culture medium in No 1 opotimized condition by gas chromatography,it showed that average fatty acid composition was C16:0 9.65%,C18:0 2.05%,C18:1n9 50.00%, C18:2n6 23.35%,C18:3n3 1.35%,C18:3n6 8.65%,C20:0 0.40%,C20:1 0.69%,C20:2n6 0.35%,C24:1(n-6) 0.60%,average contend of unsaturated fatty acids in the total fatty acids was 87.03%.
Through checking and measuring Mortierella isabellina gross oil , we discovered that other physical and chemical characters were in accord with recommended the microbial oil standard other than higher acid value.This showed that the refined strain oil was edible.
引文
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