利用伯克霍尔德氏菌脂肪酶富集裂壶藻油脂中的二十二碳六烯酸  

Concentration of Docosahexaenoic Acid in Aurantiochytrium sp. Oil by Burkholderia Lipase

作　　者：兰君[1,2] 宋晓金[1] 谭延振[1] 徐建春[3] 崔球[1,4] 冯银刚[1] 

机构地区：[1]中国科学院青岛生物能源与过程研究所青岛市单细胞油脂工程实验室山东省能源生物遗传资源重点实验室,山东青岛266101 [2]中国科学院大学生命科学学院,北京100049 [3]青岛琅琊台集团股份有限公司,山东青岛266400 [4]中国科学院青岛生物能源与过程研究所生物燃料重点实验室,山东青岛266101

出　　处：《食品科学》2015年第1期128-134,共7页Food Science

基　　金：国家高技术研究发展计划(863计划)项目(2012AA02A707;2014AA021701);国家自然科学基金青年科学基金项目(41306132)

摘　　要：目的:研究伯克霍尔德氏菌胞外脂肪酶对裂壶藻油脂中二十二碳六烯酸(docosahexaenoic acid,DHA)的富集作用。方法:用三丁酸甘油酯平板筛选法,筛选产脂肪酶的细菌;以对硝基苯月桂酸酯为底物,对筛选得到的伯克霍尔德氏菌脂肪酶的酶学性质进行研究;用该脂肪酶水解裂壶藻油脂,通过气相色谱法测定酶解后水相和有机相的脂肪酸组成来研究其对裂壶藻油脂DHA的富集作用。结果:从广州的泥土样品中分离获得一株高产胞外脂肪酶的伯克霍尔德氏菌,在产酶培养基中30℃、200 r/min的条件下培养40 h后,脂肪酶酶活力达到最大值70 U/m L。脂肪酶粗酶液酶活性的最适温度为45℃,最适p H值为8.5。该菌的脂肪酶对裂壶藻油脂中的DHA有一定的富集作用,能够使油脂中DHA占总脂肪酸的质量分数从最初的30%升高到40%。结论:筛选得到一株能够产生选择性富集DHA的脂肪酶的细菌,拓宽了可用于多不饱和脂肪酸分离纯化的脂肪酶的范围。Purpose: To explore the effect Burkholderia lipase on enrichment of docosahexaenoic acid(DHA) in Aurantiochytrium sp. oil. Methods: Tributyrin plate method was used to screen a lipase-producing bacterium. Using 4-nitrophenyl laurate as substrate, the enzymatic properties of the Burkholderia lipase were studied. The lipase was used to hydrolyze Aurantiochytrium oil. To evaluate the enrichment efficiency of DHA with the lipase, the fatty acid compositions in the aqueous and the organic phase after hydrolysis were detected by gas chromatography. Results: The lipase-producing bacterium obtained from mud samples collected from Guangzhou city was identified as Burkholderia sp. SD001. Its lipase activity reached the maximum value of 70 U/m L in the culture after fermentation at 30 ℃ for 40 h. The lipase had an optimal p H of 8.5 and an optimal temperature of 45 ℃. When the lipase was used to hydrolyze the oil from Aurantiochytrium sp., the percentage of DHA content in the oil was increased from 30% to 40%. Conclusions: A bacterium producing a lipase capable of enriching DHA has been obtained. The lipase from this strain with expanded applicability is useful for the enrichment of polyunsaturated fatty acid.

关 键 词：脂肪酶 伯克霍尔德氏菌 裂壶藻 油脂 二十二碳六烯酸富集 

分 类 号：Q939.9[生物学—微生物学] Q946