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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2023, Vol. 43 Issue (1): 115-126    DOI: 10.13523/j.cb.2207051
综述     
低温蛋白酶研究进展与应用*
郝漫,史超硕,惠威,李相勋,张彤彤,刘夫锋,路福平,张会图**()
天津科技大学生物工程学院 天津 300457
Progress and Application of Cold-active Protease
HAO Man,SHI Chao-shuo,HUI Wei,LI Xiang-xun,ZHANG Tong-tong,LIU Fu-feng,LU Fu-ping,ZHANG Hui-tu**()
College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China
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摘要：

低温蛋白酶是一类能够在低温下将大分子蛋白质水解为小分子多肽或者氨基酸的水解酶,多由低温环境中的微生物产生。低温蛋白酶在较低温度环境下具有较高的催化活性,在反应过程中仅较少的量就可以达到理想效果,并且可以避免不必要的化学转化,减少挥发性物质的损失。低温蛋白酶在洗涤、食品加工以及医疗等应用领域已经显示出独特的优势和商业价值。综述不同种类及来源的微生物低温蛋白酶,总结其催化性质、结构特征以及目前的应用情况和未来发展前景,以期进一步推动新型低温蛋白酶的开发和应用。
关键词： 低温蛋白酶;  微生物;  催化性质;  结构特征;  工业应用    
Abstract:

Cold-active protease is a kind of hydrolytic enzyme that can hydrolyze large proteins into small peptides or amino acids at low temperatures. It is generally produced by microorganisms living at low temperatures and has high catalytic activity at lower ambient temperatures. Therefore, it can avoid waste of resources caused by the rise in temperature in the application process and the special requirements for equipment, and shows unique advantages and commercial value in the application fields of washing, food processing and medical treatment. In this paper, different kinds and sources of microbial cryopinases are reviewed, and their catalytic properties, structural characteristics, current application and future development prospects are analyzed and summarized based on existing research results, aiming to promote the development and application of new cryopinases.
Key words: Cold-active protease    Microorganism    Catalytic property    Architectural feature    Industrial application
收稿日期: 2022-07-25 出版日期: 2023-02-14
ZTFLH:  Q814  
基金资助: *国家重点研发计划(2021YFC2100400)
通讯作者: **张会图 电子信箱:hzhang@tust.edu.cn   
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郝漫
史超硕
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刘夫锋
路福平
张会图

引用本文:

郝漫, 史超硕, 惠威, 李相勋, 张彤彤, 刘夫锋, 路福平, 张会图. 低温蛋白酶研究进展与应用*[J]. 中国生物工程杂志, 2023, 43(1): 115-126.

HAO Man, SHI Chao-shuo, HUI Wei, LI Xiang-xun, ZHANG Tong-tong, LIU Fu-feng, LU Fu-ping, ZHANG Hui-tu. Progress and Application of Cold-active Protease. China Biotechnology, 2023, 43(1): 115-126.

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https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2207051    或    https://manu60.magtech.com.cn/biotech/CN/Y2023/V43/I1/115

图1  低温蛋白酶研究流程
图2  产低温蛋白酶的微生物
菌种 分子量/kDa 最适作用温度/°C 最适pH 最适作用底物 参考文献
Escherichia freundii 55 25 10.0 - [6]
Pseudomonas fluorescens 47 35~40 8.0 乳蛋白、酪蛋白 [36-37]
Candida humicola 36 37 1.0~2.0 牛血清白蛋白 [33]
Vibrio sp. 35 40 8.5~9.0 - [38]
Alteromonas haloplanktis 74~76 20 8.0~9.0 - [39]
Flavobacterium balustinum 70 40 7.0~9.0 - [40]
Azospirillum sp. 48.6 40 8.5 - [41]
Leucosporidium antarcticum 34.4 25 6.7~7.1 - [42]
Flavobacterium psychrophilum 62 24 6.0~7.0 - [43]
Klebsiella oxytoca - 37 5.0~7.0 - [44]
Clostridium sp. 46 37 7.0 - [34]
Colwellia sp. 60 35 8.0~9.0 - [45]
Aspergillus ustus 32 45 9.0 - [19]
Exiguobacterium sp. 36 40 8.0 - [46]
Curtobacterium luteum 115 20 7.0 - [47]
Trichoderma atroviride 24 25 6.2 - [48]
Pseudomonas lundensis 46 30 10.4 - [49]
Serratia marcescens 56 40 8.0 - [50]
菌种 分子量/kDa 最适作用温度/°C 最适pH 最适作用底物 参考文献
Stenotrophomonas maltophilia 75 20 10.0 - [51]
Bacillus sp. - 40 7.4 - [52]
Stenotrophomonas sp. 55 15 10.0 - [53]
Acinetobacter sp. 35 40 9.0 酪蛋白、牛血清白蛋白 [54]
Halobacillus sp. 35 30 8.0 FA-Gly-Phe/Leu-NH2 [55]
Pseudoalteromonas sp. 34.6 25~35 8.0~9.0 Succ-AAPF-pNa [10]
Penicillium nalgiovense 45.2 35 8.0 - [20]
Planococcus sp. 35.6 35 10.0 - [13]
Lysobacter sp. - 40 9.0 偶氮酪蛋白 [17]
Chryseobacterium sp. - 10 7.0~8.0 酪蛋白 [16]
Planococcus sp. 43 35 10.0 - [56]
Pseudoalteromonas arctica - 30 9.0 - [57]
Bacillus sp. 62 20 9.0 - [18]
表1  低温蛋白酶的催化性质
图3  Subtilisin S41结构
低温蛋白酶 均方根偏差 模板建模评分 等效残基对 参考覆盖率/% 目标覆盖率/%
2GKO与3WHI 2.43 0.71 271 88 76
2GKO与5YL7 2.82 0.74 280 91 83
2GKO与1SH7 1.96 0.81 266 86 95
表2  低温蛋白酶结构比对分析
图4  四种低温蛋白酶结构比对结果
图5  Rosetta蛋白从头设计方法[66]
微生物 蛋白酶类型 应用领域 参考文献
Acinetobacter sp. 丝氨酸蛋白酶 洗涤剂添加剂 [54]
Arsukibacterium ikkense - 乳制品和其他功能性食品 [11]
Bacillus licheniformis 丝氨酸蛋白酶 组织解离 [75]
Bacillus sp. 金属蛋白酶 适冷性洗涤剂添加剂、环境保护、饲料添加剂 [52,81]
Bacillus subtilis 碱性蛋白酶 生物修复 [82]
Bacillus subtilis 丝氨酸蛋白酶 适冷性洗涤剂添加剂 [18]
Chryseobacterium sp. 丝氨酸蛋白酶 肉类加工 [16]
Enterococcus faecalis 金属蛋白酶 保健品 [83]
Exiguobacterium sp. - 洗涤剂添加剂 [46]
Flavobacterium limicola - 淡水沉积物矿化 [84]
微生物 蛋白酶类型 应用领域 参考文献
Pedobacter cryoconitis 金属蛋白酶 浅水生物修复 [85]
Penicillin nalgiovense 碱性蛋白酶 肉类加工 [20]
Planococcus sp. 丝氨酸蛋白酶 适冷性洗涤剂添加剂、鱼露发酵 [56,67]
Pseudomonas aeruginosa 碱性蛋白酶 洗涤 [12]
Pseudomonas sp. - 去除人发磷质层 [86]
Pseudoalteromonas sp. 丝氨酸蛋白酶 肉类嫩化、低温食品加工、皮革脱毛、海参中活性物质提取 [74,87-88]
Pseudoalteromonas arctica - 冷水洗涤 [57]
Serratia marcescens 金属蛋白酶 洗涤剂添加剂 [50]
Sphingomonas paucimobilis - 洗涤剂、食品分子生物学 [89]
Stenotrophomonas sp. 碱性蛋白酶 洗涤 [53]
表3  低温蛋白酶的应用
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