食品与发酵工业 >

2024 , Vol. 50 >Issue 10: 103 - 111

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.036282

研究报告

高效虾壳脱蛋白菌株的筛选、产酶条件优化及酶学性质研究

  • 宋超东 ,
  • 殷豆豆 ,
  • 谢晨杰 ,
  • 尹凯波 ,
  • 韦玉玲 ,
  • 李林利 ,
  • 黄宏智 ,
  • 张红岩 ,
  • 申乃坤 ,
  • 姜明国
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  • (广西民族大学 海洋与生物技术学院,广西多糖材料与改性重点实验室,广西 南宁,530006)
第一作者:硕士研究生(申乃坤教授与张红岩教授为共同通信作者,E-mail:shennaik05@126.com;hognyanzhang2008@163.com)

收稿日期: 2023-05-29

  修回日期: 2023-07-11

  网络出版日期: 2024-06-11

基金资助

国家自然科学基金项目(31660022,32060020);广西科技重点研发计划项目(AB21196019,AB21220020);广西民族大学引进人才项目(2018KJQD17);大学生创新创业训练计划课题(202110608014,S202210608164,S202210608156)

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Screening of efficient shrimp shell deproteinizing strains, optimization of enzyme production conditions and enzymatic properties

  • SONG Chaodong ,
  • YIN Doudou ,
  • XIE Chenjie ,
  • YIN Kaibo ,
  • WEI Yuling ,
  • LI Linli ,
  • HUANG Hongzhi ,
  • ZHANG Hongyan ,
  • SHEN Naikun ,
  • JIANG Mingguo
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  • (School of Marine Sciences and Biotechnology, Guangxi Minzu University, Guangxi Key Laboratory for Polysaccharide Materials and Modifications, Nanning 530006, China)

Received date: 2023-05-29

  Revised date: 2023-07-11

  Online published: 2024-06-11

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摘要

该研究旨在筛选高效虾壳脱蛋白菌株并进行工艺优化及酶学性质研究。先通过单因素、Placket-Burman(PB)和正交优化试验对筛选菌株虾壳脱蛋白工艺优化,进一步对酶学性质和降解产物进行研究。结果表明,菌株铜绿假单胞菌(Pseudomonas aeruginosa)Gxun-7脱蛋白效果最好;最优产酶条件为:虾壳粉40 g/L,玉米浆5 g/L,接种量1%(体积分数),初始pH 7.0,发酵温度35 ℃,发酵时间48 h,优化后酶活性达(617.13±22.11) U/mL,较优化前提高了43.50%,此时,虾壳脱蛋白率为(82.61±0.54)%;菌株Gxun-7所产蛋白酶为诱导酶,酶的耐盐性较好,最适温度和pH值分别为60 ℃和7.0,除Cu2+和Al3+对酶活性有显著抑制外,其他金属离子对酶活影响不大,十二烷基磺酸钠(sodium dodecyl sulfate,SDS)和十六烷基三甲基溴化铵(hexadecyl trimethyl ammonium bromide,CTAB)抑制酶活性超过40%,而β-巯基乙醇却能提高酶活性到468%;降解液中共检测出17种氨基酸,含量达1 856.14 mg/L。因此,Gxun-7对虾壳具有高效脱蛋白能力,所产蛋白酶稳定性强,降解液中氨基酸含量丰富,为虾壳蛋白降解及利用提供理论依据。

关键词： 虾壳; 脱蛋白; 铜绿假单胞菌; 条件优化; 酶学性质

本文引用格式

宋超东 , 殷豆豆 , 谢晨杰 , 尹凯波 , 韦玉玲 , 李林利 , 黄宏智 , 张红岩 , 申乃坤 , 姜明国 . 高效虾壳脱蛋白菌株的筛选、产酶条件优化及酶学性质研究[J]. 食品与发酵工业, 2024 , 50(10) : 103 -111 . DOI: 10.13995/j.cnki.11-1802/ts.036282

Abstract

This study aimed to identify exceptional deproteinization strains, and optimize their fermentation process, examine their enzymatic properties and degradation product composition.Initially, the shrimp shell deproteinization process was optimized using single-factor, Plackett-Burman (PB), and orthogonal optimization experiments.The study also analyzed the enzymatic properties of the secreted crude enzyme solution and the amino acid composition of degradation products.The result for shrimp shell protein degradation by Pseudomonas aeruginosa Gxun-7 was shrimp shell powder 40 g/L, corn pulp 5 g/L, inoculum 1% (V/V), initial pH 7.0, and the optimal fermentation temperature and time were 32 ℃ and 48 h, respectively.Under the optimized conditions, the protease activity reached (617.13±22.11) U/mL, which increased 43.50% compared with the initial strain.The percentage of deproteinized shrimp shells reached (82.61±0.54)%.The protease produced by strain Gxun-7 was an inducible enzyme, which showed high salt resistance.It had maximum activity at 60 ℃ and optimum pH at 7.0, respectively.Most metal ions had no significant effect on the protease activity, except Cu2+ and Al3+ had strong inhibitory effects.Sodium dodecyl sulfonate (SDS) and cetyltrimethylammonium bromide (CTAB) significantly reduced the enzyme activity by more than 40%, while β-mercaptoethanol increased the enzyme activity by 468%.A total of 17 amino acids were detected in the degradation solution, with concentrations up to 1 856.14 mg/L.P.aeruginosa Gxun-7 efficiently degrades proteins in shrimp shells, generating proteases with exceptional stability and abundant types and contents of amino acids in the degradation liquid.This research provides a theoretical foundation for developing shrimp shell protein fermentation technology.

Key words： shrimp shell; deproteinization; Pseudomonas aeruginosa; condition optimization; enzymatic properties

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