Marine Drugs
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2022
·Vol. 20
·期 1
·48
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IF 6.085
Jian Li,Jun-Hui Cheng,Zhao-Jie Teng,Xia Zhang,Xiu-Lan Chen,Mei-Ling Sun,Jing-Ping Wang,Yu-Zhong Zhang,Jun-Mei Ding,Xin-Min Tian,Xi-Ying Zhang
Although the S8 family in the MEROPS database contains many peptidases, only a few S8 peptidases have been applied in the preparation of bioactive oligopeptides. Bovine bone collagen is a good source for preparing collagen oligopeptides, but has been so far rarely applied in collagen peptide preparation. Here, we characterized a novel S8 gelatinase, Aa2_1884, from marine bacteriumFlocculibacter collagenilyticusSM1988T, and evaluated its potential application in the preparation of collagen oligopeptides from bovine bone collagen. Aa2_1884 is a multimodular S8 peptidase with a distinct domain architecture from other reported peptidases. The recombinant Aa2_1884 over-expressed inEscherichia colishowed high activity toward gelatin and denatured collagens, but no activity toward natural collagens, indicating that Aa2_1884 is a gelatinase. To evaluate the potential of Aa2_1884 in the preparation of collagen oligopeptides from bovine bone collagen, three enzymatic hydrolysis parameters, hydrolysis temperature, hydrolysis time and enzyme-substrate ratio (E/S), were optimized by single factor experiments, and the optimal hydrolysis conditions were determined to be reaction at 60 ℃ for 3 h with an E/S of 400 U/g. Under these conditions, the hydrolysis efficiency of bovine bone collagen by Aa2_1884 reached 95.3%. The resultant hydrolysate contained 97.8% peptides, in which peptides with a molecular weight lower than 1000 Da and 500 Da accounted for 55.1% and 39.5%, respectively, indicating that the hydrolysate was rich in oligopeptides. These results indicate that Aa2_1884 likely has a promising potential application in the preparation of collagen oligopeptide-rich hydrolysate from bovine bone collagen, which may provide a feasible way for the high-value utilization of bovine bone collagen.Keywords:peptidase;the MEROPS S8 family;bovine bone collagen;oligopeptides;hydrolysate
研究领域:
生物化学
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酶学
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食品科学
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2015
·
IF 5.636
Henan Li,Yawen Mu,Junrong Yan,Dongmei Cui,Weijun Ou,Yakun Wan,Songqin Liu
Acute renal failure (ARF) represents a very important and potentially devastating disorder in clinical nephrology. Neutrophil gelatinase-associated lipocalin (NGAL) is an early biomarker for ARF in a wide range of different disease processes, which is frequently detected in clinical diagnosis. Herein, we present a label-free and sensitive photoelectrochemical (PEC) immunosensor for NGAL by utilizing a biotinylated anti-NGAL Nanobody (Nb) orientedly immobilized to streptavidin-coated cobalt 2,9,16,23-tetraaminophthalocyanine (CoPc)-sensitized TiO2 electrode. The Nb was biotinylated at the C-terminus, which is situated at the opposite site of the antigen binding region. Using highly oriented Nb as receptor molecules, a label-free PEC immunosensor for NGAL was developed by monitoring the changes in the photocurrent signals of the electrode resulting from immunoreaction. Immobilization of Nb to streptavidin-coated CoPc-sensitized TiO2 electrode surface provides high binding capacity to NGAL; thus, it can lead to a high sensitivity. The limit of detection (LOD) of the proposed immunosensor has been significantly lowered to 0.6 pg mL–1. This proposed immunosensor reveals high specificity to detect NGAL, with acceptable intra-assay precision and excellent stability. In addition, the present work provides a new approach to design Nb-based PEC immunosensor and increases versatility of Nbs.
研究领域:
临床医学
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生物传感器
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材料科学
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DOI
PubMed
高影响因子
Foods
·
2024
·Vol. 13
·期 21
·3472
·
IF 4.7
Jia Li,Yuxin Liu,Huanhuan Yang,Luyun Cai,Wenqian Nong,Weiliang Guan
Water-free transportation (WFT) causes shrimp (Penaeus vannamei) flesh quality deterioration. However, the roles of endogenous protease-induced protein hydrolysis have been neglected in the research. In the present study, calpain zymography, gelatinase zymography, the hematoxylin–eosin staining method, and other methods were applied to investigate the response of various endogenous proteases (cathepsin, calpain, and gelatinase), the myofibril fragmentation index (MFI), and the microscopic morphology of shrimp muscle during WFT in comparison with the shrimp under the conventional water transportation strategy (WT). The results showed that the total activity of proteases in shrimp muscle increased significantly (p≤ 0.05) after simulated transportation. Cathepsins and gelatinases were activated during WFT. No significant (p> 0.05) changes of the activity of caspase-3 and the muscle cell apoptosis rate were detected in shrimp muscle cells after WFT. In addition, the MFI increased and the gap among muscle fiber bundles enlarged after WFT. Compared with WFT, no significant (p> 0.05) effect on the activities of calpain, gelatinase, and caspase-3 in the muscle of shrimp was found after WT, and only the activity of cathepsin L significantly increased (p≤ 0.05). Based on the findings, we concluded that the activation of various endogenous proteases was induced during WFT.
研究领域:
水产养殖
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蛋白质组学
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食品科学
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动物生理学
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
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2024
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IF 4.9
Yu Liu,Yawen Liu,Fangfang Wang,Zhigang Zhang,Haiming Hu,Lei Xiong,Junping Zheng,Hongtao Liu
The conventional ELISA lacked sufficient sensitivity and accuracy for analyzing neutrophil gelatinase associated lipocalin (NGAL), a potential biomarker of ulcerative colitis. Here, we developed a dual-readout plasmonic ELISA with cascade amplification of alkaline phosphatase hydrolysis and O-phenylenediamine oxidation. In this novel plasmonic ELISA, alkaline phosphatase-conjugated antibodies capture NGAL from samples, alkaline phosphatase hydrolyzes 2-phospho-L-ascorbic acid into ascorbic acid, and then ascorbic acid blocks silver-catalyzed O-phenylenediamine oxidation by turning silver ions into silver microwires. Therefore, NGAL concentration is proportional to the decrease in chromogenic and fluorescent response. This novel plasmonic ELISA exhibited a linear range of 0.5–80 ng/mL, and the LOD was as low as 0.5 ng/mL. The sensitivity was superior to that of conventional ELISA and previous studies. The plasmonic ELISA exhibited favorable selectivity because of the specific antibody-antigen interaction and the robust cascade amplification. Besides, the dual-readout feature endowed the plasmonic ELISA with advantages in terms of convenience and compatibility. In summary, our study provided a novel and sensitive plasmonic ELISA for the diagnosis of ulcerative colitis.
研究领域:
生物化学
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免疫学
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分析化学
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医学诊断
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DOI
高影响因子
Food Bioscience
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2024
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IF 4.8
Haibo Lou,Jian Wang,Yaping Wang,Yongdong Gao,Wei Wang
In this study, the probiotic characteristics of Enterococcus faecalis SN21-3 were explored by genotype and phenotype, and the safety of this strain was evaluated in vitro and vivo . The complete genome sequence of E. faecalis SN21-3 revealed basic genetic information and corresponding genes underlying its probiotic characteristics. In vitro experiments were performed with E. faecalis SN21-3, which showed prominent antimicrobial activity and excellent abilities of resistance to simulated gastrointestinal fluid, cholesterol assimilation, adhesion, and antioxidant. In addition, E. faecalis SN21-3 was susceptible to the most common antimicrobial agents, did not contain transferable resistance gene and pathogenic gene, and did not perform haemolysis reactions and gelatinase activity in vitro . Furthermore, in vivo experiments demonstrated that feeding E. faecalis SN21-3 to broilers for 28 days did not significantly affect their normal growth or blood biochemistry parameters. Moreover, H&E staining showed no evidence of lesions in the heart, liver, spleen, or kidney of the broilers. No migration of E. faecalis SN21-3 was observed in the heart, liver, spleen, kidney, or blood samples during the feeding trial, indicating that the bacteria did not cause infections. In summary, E. faecalis SN21-3 exhibits numerous probiotic characteristics and is sufficiently safe for use as a novel animal probiotic.
研究领域:
微生物学
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动物科学
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基因组学
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食品科学
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DOI
高影响因子
危险品化学品经营许可证（带存储）