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基于枯草芽孢杆菌超声辅助液态发酵豆粕的蛋白质谱分析、抗氧化活性及构效关系研究。

Analysis in protein profile, antioxidant activity and structure-activity relationship based on ultrasound-assisted liquid-state fermentation of soybean meal with Bacillus subtilis.

机构信息

School of Food and Biological Engineering, Jiangsu University, 301 Xuefu Road, Zhenjiang 212013, PR China; Institute of Food Physical Processing, Jiangsu University, 301 Xuefu Road, Zhenjiang 212013, PR China.

School of Food and Biological Engineering, Jiangsu University, 301 Xuefu Road, Zhenjiang 212013, PR China.

出版信息

Ultrason Sonochem. 2020 Jun;64:104846. doi: 10.1016/j.ultsonch.2019.104846. Epub 2019 Oct 26.

Abstract

This study investigated effects of ultrasound on the contents of peptide and soluble protein, antioxidant activity, functionalities and structural characteristics of fermented soybean meal (FSBM) with Bacillus subtilis systematically. The results showed that there were significant effects of ultrasound treatments (frequency, treatment time and power density) on the contents of peptide and soluble protein (p < 0.05). Under the optimum ultrasound conditions (power density of 0.08 W/mL, frequency of 33 kHz and treatment time of 1 h) by single factor experiment, the contents of peptide and soluble protein increased by 31.27% and 18.79% compared to those of the control, respectively. Additionally, the in vitro antioxidant activity (•OH scavenging rate, Fe chelating capacity and DPPH radical scavenging rate) and functional properties (emulsifying activity and emulsifying stability) of FSBM were found to be noticeably improved by ultrasound (p < 0.05). The fourier transform infrared (FTIR) spectroscopy revealed that ultrasound caused protein molecules to unfold with a decrease content of α-helix and β-turn and an increase in the proportion of β-sheet and random coil. Besides, atomic force microscope (AFM) results indicated that ultrasonication generally increased the surface roughness of protein and the protein sonicated with higher frequency (≥33 kHz) exhibited a greater height compared to lower frequency ultrasonication. Structure-activity relationship analysis illustrated that there was a good linear relationship between •OH scavenging rate and β-sheet/β-turn with Pearson's correlation coefficient r of -0.86/0.90. Collectively, the selection of ultrasonic parameters is essential for the preparation of functional protein and bioactive peptide by enhancing fermentation of agroindustrial by-products.

摘要

本研究系统研究了超声处理对枯草芽孢杆菌发酵豆粕(FSBM)中肽和可溶性蛋白含量、抗氧化活性、功能特性和结构特性的影响。结果表明,超声处理(频率、处理时间和功率密度)对肽和可溶性蛋白含量有显著影响(p<0.05)。在单因素实验的最佳超声条件(功率密度 0.08 W/mL、频率 33 kHz 和处理时间 1 h)下,与对照组相比,肽和可溶性蛋白含量分别提高了 31.27%和 18.79%。此外,FSBM 的体外抗氧化活性(·OH 清除率、Fe 螯合能力和 DPPH 自由基清除率)和功能特性(乳化活性和乳化稳定性)也因超声而明显改善(p<0.05)。傅里叶变换红外(FTIR)光谱表明,超声导致蛋白质分子展开,α-螺旋和β-转角含量减少,β-折叠和无规卷曲比例增加。此外,原子力显微镜(AFM)结果表明,超声普遍增加了蛋白质的表面粗糙度,且与低频超声相比,高频(≥33 kHz)超声处理的蛋白质高度更大。结构-活性关系分析表明,·OH 清除率与β-折叠/β-转角之间存在良好的线性关系,皮尔逊相关系数 r 分别为-0.86/0.90。总之,选择超声参数对于通过增强农业工业副产物的发酵来制备功能性蛋白质和生物活性肽是至关重要的。

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