食品与发酵工业 >

2023 , Vol. 49 >Issue 17: 1 - 9

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.034464

研究报告

枯草芽孢杆菌发酵紫菜制备一种α-葡萄糖苷酶抑制剂

  • 许育衔 ,
  • 程慧敏 ,
  • 韩贵新 ,
  • 毛相朝 ,
  • 姜宏
展开
  • 1(中国海洋大学 食品科学与工程学院,山东 青岛,266003)
    2(晶叶(青岛)生物科技有限公司,山东 青岛,266109)
第一作者:硕士研究生(姜宏副教授为通信作者,E-mail:jh@ouc.edu.cn)

收稿日期: 2022-11-28

  修回日期: 2023-01-20

  网络出版日期: 2023-09-27

基金资助

青岛市科技计划重点研发项目(22-3-3-hygg-28-hy);中央高校基本科研业务费项目(202262003);山东省青年科技人才托举工程项目(SDAST2021qt18)

收起

Preparation of α-glucosidase inhibitor by fermenting Porphyra with Bacillus subtilis

  • XU Yuxian ,
  • CHENG Huimin ,
  • HAN Guixin ,
  • MAO Xiangchao ,
  • JIANG Hong
Expand
  • 1(College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China)
    2(Jingye (Qingdao) Biotechnology Co.Ltd., Qingdao 266109, China)

Received date: 2022-11-28

  Revised date: 2023-01-20

  Online published: 2023-09-27

Fold

摘要

糖尿病是一种以高血糖为特征的慢性代谢疾病,由于化学降糖药物容易增加患者的肝肾负担与心血管疾病风险,因此针对轻症患者亟需一种能够控制血糖水平且低副作用的方法。许多研究表明紫菜具有降低餐后血糖的潜力,在降糖功能食品开发方面具有广阔的前景。该研究采用源自威海紫菜养殖基地的枯草芽孢杆菌(Bacillus subtilis)对条斑紫菜(Porphyra yezoensis)进行发酵,探究并优化了具有α-葡萄糖苷酶(α-glucosidase,GTase)抑制活性发酵产物的发酵条件,采用凝胶色谱法、LC-MS/MS分离并鉴定出具有GTase抑制活性的肽段,最后使用分子对接法对活性成分进行了模拟验证。结果表明,紫菜发酵粉对GTase形成竞争型抑制,IC50为0.21 mg/mL,优于阳性对照阿卡波糖(IC50=0.88 mg/mL)。成分分析与分子对接结果表明,抑制活性可能来源于肽段GPGDFL和SPPPPPA。该研究制备了一款具有GTase抑制作用的食品原料,并初步阐明了其中的主要活性成分,证明紫菜可以作为降血糖功能食品的来源之一,为紫菜的高值化开发利用提供了有效的理论支持。

关键词： 条斑紫菜; 枯草芽孢杆菌; 抗糖尿病肽; α-葡萄糖苷酶; 分子对接

本文引用格式

许育衔 , 程慧敏 , 韩贵新 , 毛相朝 , 姜宏 . 枯草芽孢杆菌发酵紫菜制备一种α-葡萄糖苷酶抑制剂[J]. 食品与发酵工业, 2023 , 49(17) : 1 -9 . DOI: 10.13995/j.cnki.11-1802/ts.034464

Abstract

Diabetes is a chronic metabolic disease characterized by high blood glucose level. Since chemical hypoglycemic drugs tend to increase the burden on the liver and kidneys and the risk of cardiovascular disease, there is an urgent need for a method to control blood glucose levels with low side effects for patients with mild disease. Some researchers have indicated that Porphyra has the capacity to decrease postprandial blood glucose levels and has potential in the development of hypoglycemic drugs and functional foods. Bacillus subtilis, isolated from a seaweed culture base in Weihai, Shandong, was used in this work to ferment Porphyra yezoensis. The fermentation conditions of the fermentation products with α-glucosidase (GTase) inhibitory activity were investigated and optimized. The potential peptides with GTase inhibitory activity were separated and identified by gel chromatography and LC-MS/MS, and the active ingredients were verified by molecular docking simulation. The results showed that P. yezoensis fermented powder (PFPOB) formed a competitive inhibition of GTase. The IC50 was 0.21 mg/mL, which was better than the positive control (Acarbose, IC50=0.88 mg/mL). Ingredient analysis and molecular docking indicated that the inhibitory activity might originate from peptides GPGDFL and SPPPPPA. In this study, a functional food ingredient with an effect on GTase inhibition was prepared, and the primary active components were first identified. This indicated that Porphyra can be one of the sources of hypoglycemic functional foods and provided theoretical support for the high-value development and utilization.

Key words： Porphyra yezoensis; Bacillus subtilis; antidiabetic peptide; α-glucosidase; molecular docking

参考文献

[1] 宋莹倩, 洪忻, 杜金玲, 等.南京市52 883名18岁及以上常住居民代谢综合征筛查结果分析[J/OL].预防医学情报杂志, 2022:1-9.(2022-11-07).https://kns.cnki.net/kcms/detail/51.1276.R.20221107.1137.002.html.
SONG Y Q, HONG X, DU J L, et al.Analysis of screening results of metabolic syndrome among 52 883 residents aged 18 years and above in Nanjing[J/OL].Journal of Preventive Medicine Information, 2022:1-9.(2022-11-07).https://kns.cnki.net/kcms/detail/51.1276.R.20221107.1137.002.html.
[2] LORENZATI B, ZUCCO C, MIGLIETTA S, et al.Oral hypoglycemic drugs:Pathophysiological basis of their mechanism of action[J].Pharmaceuticals, 2010, 3(9):3005-3020.
[3] SPILLER H A, SAWYER T S.Toxicology of oral antidiabetic medications[J].American Journal of Health-System Pharmacy, 2006, 63(10):929-938.
[4] KOJIMA Y, KIMURA T, NAKAGAWA K, et al.Effects of mulberry leaf extract rich in 1-deoxynojirimycin on blood lipid profiles in humans[J].Journal of Clinical Biochemistry and Nutrition, 2010, 47(2):155-161.
[5] 杨贤庆, 黄海潮, 潘创, 等.紫菜的营养成分、功能活性及综合利用研究进展[J].食品与发酵工业, 2020, 46(5):306-313.
YANG X Q, HUANG H C, PAN C, et al.Advances on nutrient components, biological activities and comprehensive utilization of Porphyra[J].Food and Fermentation Industries, 2020, 46(5):306-313.
[6] ZENG A Q, YANG R J, YU S H, et al.A novel hypoglycemic agent:Polysaccharides from laver (Porphyra spp.)[J].Food & Function, 2020, 11(10):9048-9056.
[7] 周锋. 蛋白酶水解条斑紫菜产活性肽的研究[D].无锡:江南大学, 2014.
ZHOU F.Study on bioactive peptide from laver hvdrolyzed by protease[D].Wuxi:Jiangnan University, 2014.
[8] 董玉婷, 蔡宏浩, 李志朋, 等.乳杆菌发酵提高坛紫菜的抗氧化和抑制糖脂代谢关键酶活性[J].食品科学, 2022, 43(10):162-171.
DONG Y T, CAI H H, LI Z P, et al.Lactobacillus fermentation improves the antioxidant activity of Porphyra haitanensis and its inhibitory activity on key enzymes of glycolipid metabolism[J].Food Science, 2022, 43(10):162-171.
[9] TAMANG J P, COTTER P D, ENDO A, et al.Fermented foods in a global age:East meets West[J].Comprehensive Reviews in Food Science and Food Safety, 2020, 19(1):184-217.
[10] ULUG S K, JAHANDIDEH F, WU J P.Novel technologies for the production of bioactive peptides[J].Trends in Food Science & Technology, 2021, 108:27-39.
[11] SANJUKTA S, RAI A K.Production of bioactive peptides during soybean fermentation and their potential health benefits[J].Trends in Food Science & Technology, 2016, 50:1-10.
[12] 尹乐斌, 李乐乐, 何平, 等.枯草芽孢杆菌发酵豆渣制备多肽及其活性研究[J].中国酿造, 2022, 41(1):75-79.
YIN L B, LI L L, HE P, et al.Preparation and activity of peptides from soybean dregs by fermentation with Bacillus subtilis[J].China Brewing, 2022, 41(1):75-79.
[13] 白露. 坛紫菜多肽的分离纯化及其抗肿瘤活性研究[D].广州:华南理工大学, 2016.
BAI L.Study on purification and anti-cancer effects of peptides derived from Porphyra haitanesis[D].Guangzhou:South China University of Technology, 2016.
[14] 董乐, 梁文文, 王芳, 等.红毛菜抗氧化肽的分离纯化及其活性评价[J].泉州师范学院学报, 2020, 38(2):9-16.
DONG L, LIANG W W, WANG F, et al.The separation and purification of antioxidant peptides from Bangia fuscopurpurea (Dillw.) Lyngb.and its activity evaluation[J].Journal of Quanzhou Normal University, 2020, 38(2):9-16.
[15] ZHAO Q, JIANG X, MAO Z J, et al.Exploration, sequence optimization and mechanism analysis of novel xanthine oxidase inhibitory peptide from Ostrea rivularis Gould[J].Food Chemistry, 2023, 404:134537.
[16] 余筱洁, 张有做, 周存山, 等.酶法水解紫菜蛋白动力学研究[J].中国食品学报, 2011, 11(3):62-67.
YU X J, ZHANG Y Z, ZHOU C S, et al.Kinetics of hydrolysis on Porphyra yezoensis protein with alkaline protease[J].Journal of Chinese Institute of Food Science and Technology, 2011, 11(3):62-67.
[17] KIERS J L, VAN LAEKEN A E A, ROMBOUTS F M, et al.In vitro digestibility of Bacillus fermented soya bean[J].International Journal of Food Microbiology, 2000, 60(2-3):163-169.
[18] 吴晖, 卓林霞, 解检清, 等.发酵条件对枯草芽孢杆菌发酵豆粕中的蛋白酶活力的影响[J].现代食品科技, 2008, 24(10):973-976.
WU H, ZHUO L X, XIE J Q, et al.Effect of fermentation conditions on the proteolytic activity of a Bacillus subtilis protease on fermented soybean meal[J].Modern Food Science and Technology, 2008, 24(10):973-976.
[19] 胡润锋, 李浚哲, 李鹏飞, 等.桑叶多糖结构特征及其对α-葡萄糖苷酶活性的抑制作用[J].林业工程学报, 2022, 7(6):100-106.
HU R F, LI J Z, LI P F, et al.Study on the structure of mulberry leaf polysaccharide and its inhibitory activity of α-glucosidase[J].Journal of Forestry Engineering, 2022, 7(6):100-106.
[20] WESTLEY A M, WESTLEY J.Enzyme inhibition in open systems[J].Journal of Biological Chemistry, 1996, 271(10):5347-5352.
[21] LIU W W, LI H Y, WEN Y Y, et al.Molecular mechanism for the α-glucosidase inhibitory effect of wheat germ peptides[J].Journal of Agricultural and Food Chemistry, 2021, 69(50):15231-15239.
[22] SIM L, QUEZADA-CALVILLO R, STERCHI E E, et al.Human intestinal maltase-glucoamylase:Crystal structure of the N-terminal catalytic subunit and basis of inhibition and substrate specificity[J].Journal of Molecular Biology, 2008, 375(3):782-792.
[23] WANG R C, ZHAO H X, PAN X X, et al.Preparation of bioactive peptides with antidiabetic, antihypertensive, and antioxidant activities and identification of α-glucosidase inhibitory peptides from soy protein[J].Food Science & Nutrition, 2019, 7(5):1848-1856.
[24] 韩芬霞, 范新景, 耿升, 等.异甘草素抑制α-葡萄糖苷酶的分子机制[J].食品科学, 2019, 40(15):37-42.
HAN F X, FAN X J, GENG S, et al.Inhibitory mechanism of isoliquiritigenin against α-glucosidase[J].Food Science, 2019, 40(15):37-42.
[25] 黄钦钦, 田亚平.条斑紫菜蛋白酶解液α-葡萄糖苷酶和DPP-IV抑制活性的表征及肽成分解析[J].食品科学, 2020, 41(24):110-116.
HUANG Q Q, TIAN Y P.α-glucosidase and dipeptidyl peptidase IV inhibitory activity and peptide composition of Porphyra yezoensis protein hydrolysate[J].Food Science, 2020, 41(24):110-116.
Options
文章导航

/

〈 〉

技术支持：北京玛格泰克科技发展有限公司