金鲳鱼内脏脂肪酶的初步纯化及鱼皮酶法脱脂的工艺优化

刘洪霞; 夏光华; 李川; 张培; 申铉日

doi:10.13386/j.issn1002-0306.2017.20.018

金鲳鱼内脏脂肪酶的初步纯化及鱼皮酶法脱脂的工艺优化

刘洪霞^1,2,
夏光华¹,
李川¹,
张培¹,
申铉日^1,2

1. 海南大学食品学院
2. 海南大学热带生物资源教育部重点实验室

基金项目:

海南省自然基金(317038)；海南省重点研发计划(ZDYF2017104)；海南省科协青年科技英才学术创新计划项目(HAST201624)；海南大学科研启动基金项目(KYQD1609)；

详细信息

作者简介:
刘洪霞 (1993-) , 女, 硕士研究生, 研究方向:水产资源高效利用技术, E-mail:lhx.jorly.26@163.com。;

申铉日 (1968-) , 男, 博士, 教授, 研究方向:热带水产品精深加工、生理活性物质的分离纯化及功能性评价、海洋医用生物材料的研究开发, E-mail:shenxuanri2009@163.com。;

中图分类号: TS254.9
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出版历程
- 收稿日期: 2017-03-03

Partial purification of lipase from golden pompano viscera and optimization of enzymatic degreasing of tilapia skin

College of Food Science and Technology, Hainan University
Key Laboratory of Tropical Biological Resource of Ministry of Education, Hainan University

摘要

摘要: 以金鲳鱼内脏为原料,采用磷酸盐缓冲液提取,硫酸铵分级沉淀的方法,初步纯化得到内脏粗脂肪酶;分别探讨了不同反应温度和p H对上述粗脂肪酶活力的影响;研究了酶在罗非鱼皮脱脂中的应用,并通过单因素实验和响应面实验优化了鱼皮酶法脱脂工艺。结果表明,经硫酸铵分级沉淀后,金鲳鱼内脏粗脂肪酶的比活力达12.60 U/mg,纯化倍数为6.06,纯度达81.65%;其最适温度为45℃,最适p H为8.0,在0~50℃及p H6.0~11.0的范围内酶活力较稳定。金鲳鱼内脏粗脂肪酶对罗非鱼皮脱脂实验结果表明,在酶添加量90 U/g,p H7.5,温度37℃,作用时间60 min的条件下,罗非鱼皮脱脂率达67.02%±1.61%,说明粗脂肪酶能在温和条件下有效脱除鱼皮脂肪。本研究为金鲳鱼加工废弃物的高值化利用提供了一定的理论依据。
- 金鲳鱼 /
- 内脏 /
- 脂肪酶 /
- 酶法脱脂
Abstract: The visceral crude lipase was extracted in phosphate buffer saline and partially purified by ammonium sulfate fractionation.The effects of different reaction temperature and p H on crude enzyme activity and stability were studied. In the application of tilapia skin degreasing, the single factor test and response surface test were used to optimize the fish skin enzymatic degreasing process. Experimental results showed that the specific activity, purification factor and purity after ( NH4) 2SO4precipitation reached 12.60 U/mg, 6.06 and 81.65%, respectively. The optimum temperature and p H was 45 ℃and 8.0, respectively.The crude lipase was relatively stable between 0⁵0 ℃ and in the p H range of 6.0¹1.0.Additionally, the crude lipase was found to be effective in degreasing of tilapia skin. The results showed that the degreasing rate of tilapia skin reached 67.02% ± 1.61% under the conditions of enzyme addition of 90 U/g, p H value of 7.5, temperature of 37 ℃ for60 min, indicating that the crude lipase could be effective in the removal of fish skin fat in mild conditions. The results offer experimental basis and theoretical reference and practical foundation for further study of the high value utilization of golden pompano processed waste.
- golden pompano /
- viscera /
- lipase /
- enzymatic degreasing

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参考文献(27)

[1]	ANTHONSEN H W, BAPTISTA A, DRABLS F, et al.Lipases and esterases:a review of their sequences, structure and evolution[J].Biotechnology Annual Review, 1995, 1 (8) :315-317.
[2]	ANDOVAL G, HC/BIOCHEMIE/BIOPHYSIK.Lipases and phospholipases:methods and protocols[M].Humana Press, 2012:3-19.
[3]	MOAYEDALLAIE S, MIRZAEI M, PATERSON J.Bread improvers:Comparison of a range of lipases with a traditional emulsifier[J].Food Chemistry, 2010, 122 (3) :495-499.
[4]	武玉婷, 李春, 王晨旭, 等.化学修饰改进脂肪酶催化特性的研究及食品中应用[J].食品工业科技, 2013, 34 (21) :366-368.
[5]	刘虹蕾, 缪铭, 江波, 等.微生物脂肪酶的研究与应用[J].食品工业科技, 2012, 33 (12) :376-381.
[6]	GALVEZ R P, BERGE J P.Utilization of fish waste:New Perspectives[M].CRC Press, 2013:7-22.
[7]	SMICHI N, GARGOURI Y, MILED N, et al.A grey mullet enzyme displaying both lipase and phospholipase activities:purification and characterization[J].International Journal of Biological Macromolecules, 2013, 58:87-94.
[8]	SMICHI N, FENDRI A, CHABOUNI R, et al.Purification and biochemical characterization of an acid-stable lipase from the pyloric caeca of sardine (Sardinella aurita) [J].Applied Biochemistry and Biotechnology, 2010, 162 (5) :1483-1496.
[9]	KURTOVIC I, MARSHALL S N, ZHAO X, et al.Purification and properties of digestive lipases from Chinook salmon (Oncorhynchus tshawytscha) and New Zealand hoki (Macruronus novaezelandiae) [J].Fish Physiology and Biochemistry, 2010, 36 (4) :1041-1060.
[10]	戴梓茹, 钟秋平, 林美芳, 等.金鲳鱼营养成分分析与评价[J].食品工业科技, 2013, 34 (1) :347-350.
[11]	孙立元, 郭华阳, 朱彩艳, 等.卵形鲳鲹育种群体遗传多样性分析[J].南方水产科学, 2014, 10 (2) :67-71.
[12]	陈胜军, 李来好.罗非鱼加工产业相关情况[J].科学养鱼, 2014 (9) :16-17.
[13]	李佳, 许永安.罗非鱼鱼皮提取明胶的酶法脱脂工艺研究[J].福建水产, 2010 (3) :26-30.
[14]	张立坚, 杨会邦, 张俊杰, 等.罗非鱼不同组织脂肪酸含量的分析[J].淡水渔业, 2010, 40 (2) :36-40.
[15]	陈胜军, 李来好, 杨贤庆, 等.脂肪酶在鲻鱼脱脂中的应用[J].食品科学, 2007, 28 (2) :153-155.
[16]	蒋丽, 陆剑锋, 吕顺, 等.响应面法优化罗非鱼皮酶法脱脂工艺[J].食品工业科技, 2016, 37 (6) :211-215.
[17]	吴燕燕, 朱小静.鱼类脱脂方法研究进展[J].核农学报, 2016, 30 (6) :1163-1168.
[18]	SMICHI N, FENDRI A, GARGOURI Y, et al.A high salttolerant thermoactive esterase from golden grey mullet:purification, characterization and kinetic properties[J].Journal of Food Biochemistry, 2015, 39 (3) :289-299.
[19]	WINKLER U K, STUCKMANN M.Glycogen, hyaluronate, and some other polysaccharides greatly enhance the formation of exolipase by Serratia marcescens[J].Journal of Bacteriology, 1979, 138 (3) :663-670.
[20]	BRADFORD M M.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J].Analytical Biochemistry, 1976, 72 (1-2) :248-254.
[21]	LAEMMLI U K.Cleavage of structural proteins during the assembly of the head of bacteriophage T4[J].Nature, 1970, 227:680-685.
[22]	NORIEGA-RODRíGUEZ J A, GáMEZ-MEZA N, MEDINA-JUáREZ L A, et al.Extraction and fractionation of lipolytic enzyme from viscera of Monterey sardine (Sardinops sagax caerulea) [J].International Journal of Food Science&Technology, 2009, 44 (6) :1223-1228.
[23]	CHERIF S, FENDRI A, MILED N, et al.Crab digestive lipase acting at high temperature:purification and biochemical characterization[J].Biochimie, 2007, 89 (8) :1012-1018.
[24]	张培, 申铉日, 李川, 等.金鲳鱼内脏酸性蛋白酶的分离纯化及酶学性质研究[J].食品工业科技, 2017 (2) :210-214.
[25]	梁鹏, 赵卉双, 安然, 等.碱性脂肪酶对鲶鱼鱼皮脱脂效果的影响[J].食品科技, 2015 (6) :147-150.
[26]	曾小峰, 白小鸣, 盖智星, 等.响应面实验优化超声辅助提取柚皮纤维素工艺[J].食品科学, 2015, 36 (14) :34-38.
[27]	吴祥庭.酶法提油技术的研究进展[J].粮油食品科技, 2006, 14 (6) :41-42.

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