|本期目录/Table of Contents|

[1]屈家亮,柳惠平,张佑红*.纳豆激酶研究进展[J].武汉工程大学学报,2024,46(05):527-533.[doi:10.19843/j.cnki.CN42-1779/TQ.202402008]
 QU Jialiang,LIU Huiping,ZHANG Youhong*.Review on nattokinase research[J].Journal of Wuhan Institute of Technology,2024,46(05):527-533.[doi:10.19843/j.cnki.CN42-1779/TQ.202402008]
点击复制

纳豆激酶研究进展(/HTML)
分享到:

《武汉工程大学学报》[ISSN:1674-2869/CN:42-1779/TQ]

卷:
46
期数:
2024年05期
页码:
527-533
栏目:
生物与环境工程
出版日期:
2024-10-28

文章信息/Info

Title:
Review on nattokinase research
文章编号:
1674 - 2869(2024)05 - 0527 - 07
作者:
屈家亮1柳惠平2张佑红*1
1. 武汉工程大学环境生态与生物工程学院,湖北 武汉 430205;
2. 武汉工程大学光信息与能源学院和数理学院,湖北 武汉 430205
Author(s):
QU Jialiang1LIU Huiping2ZHANG Youhong*1
1. School of Environmental Ecology and Biological Engineering, Wuhan Institute of Technology,Wuhan 430205,China;
2. School of Optical Information and Energy Engineering, School of Mathematics and Physics, Wuhan Institute of Technology, Wuhan 430205, China
关键词:
纳豆激酶溶栓理化特性进展
Keywords:
nattokinase thrombolysis physicochemical properties review
分类号:
TS201
DOI:
10.19843/j.cnki.CN42-1779/TQ.202402008
文献标志码:
A
摘要:
纳豆激酶(NK)主要是由枯草芽孢杆菌代谢产生的一种碱性丝氨酸蛋白酶,具有很好的溶栓效应,有望成为新一代溶栓药物。首先从NK的基因组成及信号肽、前导肽和成熟肽的结构和功能等方面阐述了对NK的分子生物学研究。其次从酶的温度、pH、离子影响和最适底物类型等酶学性质对NK的特性进行了总结。再者对NK的三级结构及催化位点进行了分析,以位于活性中心的催化三联体(Asp32,His64,Ser221)为例探讨了NK与底物结合及催化的机制。再根据已有的文献报道,从NK的菌种改造、分离纯化及NK不同方式制备的剂型等方面介绍了国内外目前的研究现状。最后,在菌种改造和分离纯化等研究方向上对NK的未来研究进行了分析和展望,如可以使用基因编辑技术对菌种的代谢通路进行改造,达到提高产量的目的;采用分子印迹技术纯化NK。
Abstract:
Nattokinase is a type of alkaline serine protease primarily produced by the metabolism of Bacillus subtilis. It exhibits excellent thrombolytic effects and holds promise as a new generation of thrombolytic drugs. We start by explaining the molecular biology of nattokinase, including its gene composition, signal peptide, leader peptide, and the structure and function of the mature peptide. Then, we summarize the enzymatic properties of nattokinase, such as temperature, pH, ionic effects, and the best substrate type. Moreover, we analyze the tertiary structure and catalytic sites of nattokinase, and take the catalytic triad (Asp32, His64, Ser221) in the active center as an example to study the mechanism of nattokinase-substrate binding and catalysis. In addition, based on the existing literature, we give an overview of the current global research situation of nattokinase, including strain modification, isolation, purification, and formulation of various preparations. Finally, in the context of strain modification, isolation and purification, we make an analysis and prediction of the future research direction of nattokinase. For example, using gene editing techniques to engineer the metabolic pathways of the strain may increase the yield; using molecular imprinting technology for purification of nattokinase has great potential.

参考文献/References:

[1] 《中国心血管健康与疾病报告2021》编写组. 《中国心血管健康与疾病报告2021》要点解读 [J].中国心血管杂志, 2022, 27 (4) : 305-318.

[2] HONGJIE C, MCGOWAN E, REN N,et al. Nattokinase: a promising alternative in prevention and treatment of cardiovascular diseases [C]. Apsa-ascept Joint Scientific Meeting. 2018.
[3] SUMI H, HAMADA H, TSUSHIMA H, et al. A novel fibrinolytic enzyme (nattokinase) in the vegetable cheese Natto; a typical and popular soybean food in the Japanese diet [J]. Experientia, 1987, 43(10): 1110.
[4] 孟繁宇, 薛菲, 施慧. 纳豆激酶对动脉粥样硬化模型大鼠血脂及血液流变学影响 [J]. 中国实验诊断学, 2013, 17(9): 1567-1569.
[5] GITTE S J, MIKI L, ERO M P, et al. Consumption of nattokinase is associated with reduced blood pressure and von willebrand factor, a cardiovascular risk marker: results from a randomized, double-blind, placebo-controlled, multicenter North American clinical trial [J]. Integrated Blood Pressure Control, 2016, 9:95-104.
[6] 占慧琴, 袁志兰, 杨翎, 等. 纳豆激酶对视网膜中央静脉阻塞动物模型中闪光视网膜电图的影响 [J]. 南京医科大学学报(自然科学版),2010,30 (7):974-976.
[7] 高泽鑫, 何腊平, 刘亚兵, 等. 纳豆激酶的研究进展与展望 [J].中国酿造,2017,36 (8):11-15.
[8] SUZUKI Y, KONDO K, IEHISE J, et al. Dietary supplementation with fermented soybeans suppresses intimal thickening[J]. Nutrition,2003,19(3):261-264.
[9] NAKAMURA T, YAMAGATA Y, ICHISHIMA E. Nucleotide sequence of the subtilisin NAT gene, aprN, of Bacillus subtilis (Natto) [J]. Bioscience, Biotechnology, and Biochemistry,1992,56(11): 1869-1871.
[10] JIA Y, LIU H, BAO W, et al. Functional analysis of propeptide as an intramolecular chaperone for in vivo folding of subtilisin Nattokinase [J]. FEBS Letters,2010, 584(23) : 4789-4796.
[11] 贾焱. 纳豆激酶分子内分子伴侣的研究 [D]. 武汉: 武汉大学, 2013.
[12] URANO T. The profibrinolytic enzyme subtilisin NAT purified from Bacillus subtilis cleaves and inactivates plasminogen activator inhibitor type 1 [J]. Journal of Biological Chemistry, 2001, 276 (27) : 24690-24696.
[13] 于潇潇, 杜慧聪, 商鹤琴, 等. 纳豆激酶活性和稳定性的研究进展 [J]. 中国调味品, 2019, 44 (7) : 160-163.
[14] YANAGISAWA Y, CHATAKE T, CHIBA-KAMOSHIDA K. et al. Purification, crystallization and preliminary X-ray diffraction experiment of Nattokinase from Bacillus subtilis Natto [J]. Acta Crystallographica Section F,2010, 66(12) : 1670-1673.
[15] CARTER P, WELLS J A. Dissecting the catalytic triad of a serine protease [J]. Nature, 1988, 332(7): 564-568.
[16] 杨亚楠, 陈慧洁, 余河水, 等. 纳豆激酶稳定性研究 [J]. 辽宁中医药大学学报, 2018,20 (11):41-44.
[17] 李睿, 阮文辉, 熊晓辉. 纳豆激酶的酶学性质研究 [J]. 扬州大学学报(农业与生命科学版), 2015,36 (2) : 106-110.
[18] SUMI H. A novel fibrinolytic enzyme in the vegetable Natto: a typical and popular soybean food in the Japanese diet [J]. Expvrience,1987, 43 (20) : 1110-1111.
[19] 孙江伟, 王莹莹, 李琳, 等. 纳豆激酶及其药理作用的研究进展 [J]. 中国现代医药杂志,2011,13 (9) : 131-133.
[20] ICHISHIMA E, TAKADA Y, TAIRA K, et al. Specificities of extracellular and ribosomal serine proteinases from Bacillus Natto, a food microorganism [J]. Biochimica Et Biophysica Acta, 1986, 869 (2):178-184.
[21] YIN L J, LIN H H, JIANG S T, et al. Bioproperties of potent Nattokinase from Bacillus subtilis YJ1 [J]. Journal of Agricultural and Food Chemistry, 2010, 58(9): 5737-5742.
[22] 林飞龙, 李泰明. 纳豆激酶及其生物工程研究进展 [J]. 生物医学, 2023, 13 (2):240-246.
[23] LI Y,CHEN L Q,TANG X Y,et al. Biotechnology, bioengineering and applications of Bacillus Nattokinase [J]. Biomolecules, 2022,12 (7):980.
[24] YATAGAI C, MARUYAMA M, KAWAHARA T, et al. Nattokinase-promoted tissue plasminogen activator release from human cells [J]. Pathophysiology of Haemostasis and Thrombosis,2007,36(5):227-232.
[25] KUNAMNENI A, DURVASULA R. Streptokinase: a drug for thrombolytic therapy: a patent review [J]. Recent Adv Cardiovascular Drug Discovery, 2014, 9 (2):106-121.
[26] AMIN K, ZENG X B, YING Y, et al. Effect of pulsed electric fields(PEF) on protease and antioxidant activity of selenium modified fibrinolytic enzyme Nattokinase produced by Bacillus nattocells [J]. International Journal of Biosciences,2019,14 (5):282-293.
[27] HASSANNEJAD S, RAZEGHIAN E, SHOJAEI N. Relationship between PAI1 promoter 4G/5g polymorphism and stroke [J]. Basic & Clinical Medicine, 2011, 5 (17) : 18-20.
[28] 闫泉香, 冯利, 徐峰, 等. 纳豆激酶的溶栓作用及其机制研究 [J]. 食品工业科技, 2021, 42 (24) : 340-347.
[29] 杨子琼, 李迪文, 赵小斌, 等. 纳豆激酶高产菌株的复合诱变选育 [J]. 武汉工程大学学报, 2022,44(1) : 76-80.
[30] 巩涛, 魏传军, 安明理, 等. 1株纳豆激酶高产菌株的分离筛选与复合诱变研究 [J]. 安徽农业科学, 2021, 49 (15): 159-161.
[31] WANG Y X, WANG J, ZHANG X A, et al. Genomic and transcriptomic analysis of Bacillus subtilis JNFE1126 with higher Nattokinase production through ultraviolet combined Co-60-gamma ray mutagenesis [J]. Lebensmittel-Wissenschaft & Technologie,2021,147 (2):111652- 111662.
[32] CAI Y J, WEI B, JIANG S J, et al. Directed evolution improves the fibrinolytic activity of Nattokinase from Bacillus Natto [J]. FEMS Microbiology Letters, 2011, 325 (2): 155-161.
[33] CAI D B, WEI X T, QIU Y M, et al. High-Level expression of Nattokinase in Bacillus licheniformis by manipulating signal peptide and signal peptidase [J]. Journal of Applied Microbiology,2016,121(3):704-712.
[34] 刘中美,何孝天,崔文璟,等. 通过定点突变增强纳豆激酶的热稳定性 [J]. 现代食品科技, 2015,31 (2):37-41.
[35] 刘朔, 姜梅, 陈晓红, 等. 纳豆激酶第36位氨基酸突变对其活性及热稳定性的影响 [J]. 南京农业大学学报,2008 (3):130-136.
[36] 王刚, 郭明珠, 陈光. 枯草芽孢杆菌纳豆激酶分离纯化及酶学性质 [J].食品科学, 2012, 33 (15) : 231-234.
[37] 史延茂, 马跃华, 杨明, 等. 纳豆激酶分离纯化方法的研究 [J]. 中国酿造, 2010(4):119-123.
[38] 李睿, 阮文辉. 纳豆激酶NKⅡ分离纯化及其酶促动力学研究 [J]. 中国酿造, 2016, 35 (7):89-92.
[39] YAN J K, WANG Y Y, WEN Y Q, et al. Three phase partitioning as an elegant and versatile platform applied to nonchromatographic bioseparation processes [J]. Critical Reviews in Food Science & Nutrition, 2017, 58 (14) : 2416-2431.
[40] GAGAOUA M, HAFID K. Three phase partitioning system, an emerging non-chromatographic tool for proteolytic enzymes recovery and purification [J]. Biosensors Journal,2016,5 (1):100134.
[41] GARG R, THORAT B N. Nattokinase purification by three phase partitioning and impact of t-butanol on freeze drying [J]. Separation & Purification Technology, 2014, 131: 19-26.
[42] 刘俊果, 邢建民, 畅天狮, 等. 反胶团萃取分离纯化纳豆激酶 [J]. 科学通报,2006(2):133-137.
[43] 胡洪波,张雪洪,梅乐和,等. 膨胀床离子交换吸附分离纳豆激酶 [J]. 化学工程, 2005(4):1-4.
[44] YANG C L, XING J M, GUAN Y P, et al. Superparamagnetic poly (methyl methacrylate) beads for Nattokinase purification from fermentation broth [J]. Applied Microbiology and Biotechnology, 2006, 72 (3):616-622.
[45] SONG R Y, HU X L, QIAN L W, et al. Synthesis of glutathione imprinted polymer particles via controlled living radical precipitation polymerization[J]. Chinese Journal of Polymer Science,2015,33(3): 404-415.
[46] 张勋力, 张迎庆. 纳豆激酶表面分子印迹聚合物及其制备方法和应用:CN201910742958.0 [P]. 2024-01-08.
[47] 马晓文, 肖菊香, 莫炜, 等. 超临界CO2流体技术制备纳豆激酶肠溶微丸的初探 [J]. 药物生物技术, 2008,15 (3):199-203.
[48] 邓柳艳, 张建华. 纳豆激酶微胶囊制备及其释放与CaCo2细胞模型吸收的评价 [J]. 食品工业科技, 2019, 40 (10) : 115-127.
[49] 张佑红,李迪文.一种纳豆激酶肠溶胶囊及制备方法: CN202210825719.3 [P]. 2023-11-28.
[50] WANG X N, JIANG S F, WANG X Y, et al. Preparation and evaluation of Nattokinase-loaded self-double-emulsifying drug delivery system [J]. Asian Journal of Pharmaceutical Sciences. 2015, 10 (5) : 386-395.
[51] DONG X Y, KONG F P, YUAN G Y, et al. Optimisation of preparation conditions and properties of phytosterol liposome-encapsulating Nattokinase [J]. Natural Product Research,2012,26 (6):548-556.
[52] 杨超, 尹宗宁, 俞功龙, 等. 纳豆激酶缓释片的制备及药物释放影响因素的研究 [J]. 中国新药杂志, 2007,16 (7):547-551.
[53] FENG R, LI J, CHEN J P, et al. Preparation and toxicity evaluation of a novel Nattokinase-tauroursodeoxycholate complex [J]. Asian Journal of Pharmaceutical Sciences,2018,13 (2):173-182.

相似文献/References:

备注/Memo

备注/Memo:
收稿日期:2024-02-27
基金项目:武汉工程大学第十四届研究生教育创新基金(CX2022555)
作者简介:屈家亮, 硕士研究生, Email: [email protected]
*通信作者:张佑红,博士,教授。 Email: [email protected]
引文格式:屈家亮,柳惠平,张佑红. 纳豆激酶研究进展[J]. 武汉工程大学学报,2024,46(5):527-533.
更新日期/Last Update: 2024-10-26