解脂耶氏酵母lip2脂肪酶水解谷维素产生阿魏酸的研究

张宸; 严勃; 孟永宏; 郭玉蓉; 邓红; 魏丽娜

doi:10.13386/j.issn1002-0306.2014.22.033

解脂耶氏酵母lip2脂肪酶水解谷维素产生阿魏酸的研究

陕西师范大学食品工程与营养科学学院

基金项目:

陕西省科技统筹(2011KTCQ02-04)；教育部博士点专项科研基金(20130202120007)； 2012年度中央高校科研基本业务费重点项目；

详细信息

作者简介:
张宸 (1988-) , 女, 硕士研究生, 主要从事食品生物工程方面的研究。;

孟永宏 (1975-) , 男, 博士, 副教授, 主要从事食品生物工程方面的研究。;

中图分类号: TQ929;TQ460.6
计量
- 文章访问数: 0204
- HTML全文浏览量: 033
- PDF下载量: 0177
出版历程
- 收稿日期: 2014-02-23

Production of ferulic acid from oryzanol by Y. lipolytica lip2 enzyme-catalyzed

Shaanxi Normal University, College of Food Engineering and Nutritional Science

摘要

摘要: 对解脂耶氏酵母（Yarrowia lipolytica）脂肪酶水解谷维素产生阿魏酸的酶反应体系进行了研究。实验发现解脂耶氏酵母全脂肪酶粉（105U/mg）在50mmol/L p H7.0 Tris-HCl（含7.5mmol/L黄胆酸钠）,100mmol/L p H6.0磷酸钠缓冲液（含1000U脂肪酶）的体系中,水解产生阿魏酸的得率为2.94%。为了进一步提高脂肪酶水解效率,对解脂耶氏酵母脂肪酶中主要组分lip2脂肪酶基因进行了克隆,整合至毕赤酵母GS115基因组后发酵制取lip2脂肪酶粉（70.1U/mg）,于上述酶解体系中进行水解谷维素实验。实验结果表明阿魏酸产率为2.87%。获得的lip2脂肪酶催化效率略低于全脂肪酶粉催化效率,但是获得了单一的脂肪酶基因,为进一步采取分子进化技术提高其催化能力奠定了基础。
- 阿魏酸 /
- 脂肪酶 /
- 解脂耶氏酵母 /
- 酶解工艺
Abstract: An appropriate enzymatic system was preliminary explored for the Yarrowia lipolytica lipase. The study found that the optimal enzyme reaction system for the whole lipase powder was 50mmol/L p H7.0 Tris-HCl (containing 7.5mmol/L ammonium taurocholate) , 100mmol/L p H6.0 PBS (containing 1000 U lipase powder) , and the ferulic acid yield was 2.94%. To further improve the efficiency of hydrolysis of lipase, lip2 lipase gene was cloned and integrated in Pichia pastoris GS115 genome, and the lip2 lipase powder was obtained by fermentation. Using the optimal enzyme reaction system to hydrolysis oryzanol, the results showed that the ferulic acid yield by lip2 lipase powder was 2.87%. This efficiency was slightly lower than the whole lipase powder hydrolysis efficiency. However, a single lipase gene was obtained, which would lay the foundation of improving the catalyze ability to hydrolysis oryzanol by molecular evolution technology.
- ferulic acid /
- lipase /
- Yarrowia lipolytica /
- hydrolysis process

HTML全文

参考文献(19)

[1]	梁娜, 孙少平, 罗跃娥, 等.阿魏酸的研究进展[J].黑龙江中医药, 2009 (3) :39-40.
[2]	Ou Shiyi, Kwok K C.Review Ferulic acid:pharmaceutical functions, preparation and applications in foods[J].J Sci Food Agr, 2004, 84 (11) :1261-1269.
[3]	邵艳清, 李海燕.阿魏酸含量分析方法概述[J].中华中医药杂志, 2009, 24 (4) :505-506.
[4]	赵东平, 杨文钰, 陈兴福.阿魏酸的研究进展[J].时珍国医国药, 2008, 19 (8) :1839-1841.
[5]	张璟, 欧仕益, 张宁, 等.酶解麦麸制备阿魏酸和低聚糖的研究[J].食品科学, 2004, 24 (11) :63-68.
[6]	Sancho A I, BartoloméB, Gómez-Cordovés, et al.Release of ferulic acid fromcereal residues by barley enzymatic extract[J].J Cereal Sci, 2001, 34 (2) :173-179.
[7]	Kroon P A, Garcia-Conesa M T, Fillingham I J, et al.Release of ferulic acid dchydrodimers from plant cellwalls by feruloyl esterases[J].J Sci Food Agric, 1999, 79:428-434.
[8]	Miller A, Majauskaite L, Engel K H.Enzyme-catalyzed hydrolysis ofγ-oryzanol[J].Eur Food Res Technol, 2004, 218 (4) :349-354.
[9]	Mathew S, Abraham T E.Studies on the production of feruloyl esterase from cereal brans and sugar cane bagasse by microbial fermentation[J].Enzyme Microb Technol, 2005, 36 (4) :565-570.
[10]	Madzak C, Gaillardin C, Beckerich J M.Heterologous protein expression and secretion in the non-conventional yeast Yarrowia lipolytica:a review[J].J Biotechnol, 2004, 109 (1) :63-81.
[11]	Yu M, Wen S, Tan T.Enhancing production of Yarrowia lipolytica lipase lip2 in Pichia pastoris[J].Eng Life Sci, 2010, 10 (5) :458-464.
[12]	Rywińska A, Juszczyk P, Wojtatowicz M, et al.Glycerol as a promising substrate for Yarrowia lipolytica biotechnological applications[J].Biomass Bioenergy, 2013, 48:148-166.
[13]	李璟, 王燕, 杨山虎, 等.固定化微生物脂肪酶粉转化生物柴油研究[J].浙江农业学报, 2012, 24 (3) :494-498.
[14]	杨华, 娄永江.国产碱性脂肪酶的测定方法及特性研究[J].中国食品学报, 2006, 6 (3) :138-142.
[15]	廖律, 周明达, 肖劲, 等.米糠中阿魏酸的提取及高效液相色谱法测定[J].食品工业科技, 2007, 28 (2) :217-219.
[16]	Laura N, Robert A M, Lamp A M, et al.Enzymatic hydrolysis of steryl ferulates and steryl glycosides[J].Eur Food Res Technol, 2008, 227 (3) :727-733.
[17]	Mc Clendon S D, Shin H D, Chen R R.Novel bacterial ferulic acid esterase from Cellvibrio japonicus and its application in ferulic acid release and xylan hydrolysis[J].Biotechnol Lett, 2011, 33:47-54.
[18]	郭美锦, 朱泰承, 张明, 等.重组毕赤酵母甲醇利用表型与基因拷贝数对外源基因表达的影响[J].中国生物工程杂志, 2007, 27 (7) :7-11.
[19]	Lanser A C, Manthey L K, Hou C T.Regioselectivity of new bacterial lipases determined by hydrolysis of triolein[J].Curr Microbiol, 2002, 44 (5) :336-340.