JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
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2021
·Vol. 101
·期 13
·5645-5651
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IF 3.639
Linzi Yang,Xueli Li,Yunhao Lu,Yuanping Lv,Yuanlong Chi,Qiang He
BACKGROUND Esters are indispensable aroma compounds and contribute significantly to the fruity aromas in fermented condiments. The ester synthesis activity and pathways of Bacillus licheniformis , Candida etchellsii, and Zygosaccharomyces rouxii, isolated from Chinese horse bean chili-paste (CHCP), were investigated. Chemical buffer models containing esterification and alcoholysis systems inoculated with extracellular extracts of these three strains were established. RESULTS The ester synthesis activity of C. etchellsii was stronger than that of the other two strains. Zygosaccharomyces rouxii could synthesize acetate esters via esterification, whereas the biosynthesis pathways of B. licheniformis and C. etchellsii were esterification and alcoholysis. Esterification exhibited relatively high activity at pH 4, whereas alcoholysis activity improved with an increase in the pH from 4 to 8. Candida etchellsii could synthesize C 6 –C 8 of acetate esters, and its activity improved with the number of alcohol carbon atoms. These three strains could synthesize C 10 –C 18 of ethyl esters. Their ethyl ester synthesis activity decreased with the aliphatic acid carbon number. CONCLUSION Candida etchellsii has the potential to be used in CHCP fermentation to accumulate esters and improve flavor compared with the other two strains. This research is helpful in explaining the mechanism of ester synthesis in fermented condiments. © 2021 Society of Chemical Industry.
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ANALYST
·
2020
·Vol. 145
·期 9
·3257-3265
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IF 3.978
Yue Li,Xinxin Xu,Liqiang Liu,Hua Kuang,Liguang Xu,Chuanlai Xu
In this study, a gold immunochromatographic assay (GICA) based on a penicillin receptor protein (PBP) is proposed to simultaneously detect penicillin, cephalosporin, and carbapenem antibiotics in milk and chicken. A mutant PBP, carboxy-terminal of the β-lactam sensor-transducer protein (BlaR-CTD, I188KS19CG24C) from Bacillus licheniformis ATCC14580 was expressed in E. coli as a soluble His-tag fusion protein. The purified BlaR-CTD mutant was labeled with colloidal gold (CG-BlaR-CTD-M) and applied to develop GICA. The GICA method was performed based on a competitive form, where the binding of the ampicillin-bovine serum albumin conjugate (Amp-BSA) to CG-BlaR-CTD-M was inhibited by free β-lactams in samples. The performance of the assay was confirmed by analysis of milk and chicken samples. The limits of detection of 21 β-lactam antibiotics met the maximum residue limits set by the European Union and China. The GICA gave results within 10 min, thus providing a rapid and reliable method for the on-site detection of multiple β-lactams in milk and chicken samples.
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BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
·
2019
·Vol. 67
·期 2
·249-256
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IF 1.559
Bingfeng Li,Bo Fan,Jingping Fan,Siyuan Chang,Xin Pan,Yinan Wang,Yaming Wu,Junsong Song,Xuejun He
Raspberry ketone is a primary aroma component of the red raspberry. The glycosylation of this compound is a potential approach used to improve its pharmaceutical properties. In this work, raspberry ketone glycosides are produced in bacteria for the first time. Bacillus licheniformis PI15, an organic solvent-tolerant glycosyltransferase-producing strain, was isolated from chemically polluted soil. The cloning and heterologous expression of a glycosyltransferase, which was designated PI-GT1, in Escherichia coli BL21 resulted in the expression of an active and soluble protein that accounted for 15% of the total cell protein content. Purified PI-GT1 was highly active and stable over a broad pH range (6.0–10.0) and showed excellent pH stability. PI-GT1 maintained almost 60% of its maximal activity after 3 H of incubation at 20–40 °C and demonstrated optimal activity at 30 °C. Additionally, PI-GT1 displayed high stability and activity in the presence of hydrophilic solvents with log P ≤ −0.2 and retained more than 80% of its activity after 3 H of treatment. Supplementation with 10% DMSO markedly improved the glycosylation of raspberry ketone, resulting in a value 26 times higher than that in aqueous solution. The organic solvent-tolerant PI-GT1 may have potential uses in industrial chemical and pharmaceutical synthesis applications.
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JOURNAL OF SEPARATION SCIENCE
·
2017
·Vol. 40
·期 22
·4446-4452
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IF 2.557
Wei Wang,Ruiguang Bai,Xiaoyu Cai,Ping Lin,Lihong Ma
A method using high-speed capillary micellar electrokinetic chromatography and a microbial fuel cell was applied to determine the metabolite of the peptides released by Bacillus licheniformis . Two peptides, l -carnosine and l -alanyl- l -glutamine were used as the substrate to feed Bacillus licheniformis in a microbial fuel cell. The metabolism process of the bacterium was monitored by analyzing the voltage outputs of the microbial fuel cell. A home-made spontaneous injection device was applied to perform high-speed capillary micellar electrokinetic chromatography. Under the optimized conditions, tryptophan, glycine, valine, tyrosine and the two peptides could be rapidly separated within 2.5 min with micellar electrokinetic chromatography mode. Then the method was applied to analyze the solutions sampled from the microbial fuel cell. After 92 h running, valine, as the metabolite, was successfully detected with concentration 3.90 × 10 −5 M. The results demonstrated that Bacillus licheniformis could convert l -carnosine and l -alanyl- l -glutamine into valine. The method employed in this work was proved to have great potential in analysis of metabolites, such as amino acids, for microorganisms.
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BMC MICROBIOLOGY
·
2025
·Vol. 25
·期 1
·1-19
·
IF 4.2
Xu Zeyan,Zhao Lijiang,Wu Mengting,Cui Anqi,Chen Wei,Zheng Guohao,Zhou Jingyi,Gao Daqing,Shi Ruihua
Background Inflammatory bowel disease (IBD) involves the complex interplay among the mucosal barrier, microbiota, immunity and genetic factors. There are currently no satisfactory treatments for IBD. Administration of the probiotic Bacillus licheniformis (Bl) can improves colitis by regulating the gut microbiota. The phytohormone abscisic acid (ABA) treatment has favorable effects on immunity, as well as on inflammatory diseases like colitis. We hypothesized that the expression of an additional cyp gene by the Bl to increase its ABA production would enhance its effects. Results In this study, we found that a Bl-cyp strain overexpressing the cyp gene secreted more ABA into its supernatant than either the parental Bl stain or a Bl-pET82a strain expressing only a vector pET82a when these bacteria were grown in Nfb medium for 48 h. The prophylactic administration of the Bl-cyp strain culture more effectively attenuated dextran sodium sulfate (DSS)-induced colitis in mice compared to the Bl and Bl-pET28a strains. These findings were associated with significantly reduced epithelial barrier damage, as well as increased number of goblet cells and expression levels of occludin gene in the colonic epithelial layer, and decreased serum LPS levels in the Bl-cyp group. In addition, the administration of Bl-cyp strain effectively regulated the disordered gut microbiota by improving their diversity, richness and compositions more than the Bl or Bl-pET82a strain, including the ratio of Bacteroidota : Bacillota . It also inhibited the excessive growth of opportunistic pathogen Escherichia just like the Bl or Bl-pET82a strain. Moreover, the preventive administration of the Bl-cyp strain to mice following DSS-induced colitis enhanced the proportion of Treg cells and suppressed the proportion of Th17 cells in mesenteric lymph nodes (MLNs), decreased the levels of pro-inflammatory cytokines TNF-α, IL-6, and IL-22, and increased the level of anti-inflammatory IL-10 in colon tissues, similar to treatment with a high concentration of the ABA standard (ABA-H). Notably, the treatment with the Bl-cyp strain more effectively regulated the disordered microbiota than the ABA-H. Conclusions The administration of the Bl-cyp strain may provide a novel preventive approach for IBD, and may exert its effects by modulating the gut microbiota and host’s immune status.
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2023
·
IF 8.2
Yawen Zhou,Juan Shen,Huibing Chi,Xiaoyu Zhu,Zhaoxin Lu,Fengxia Lu,Ping Zhu
Type II L-asparaginase (ASNase) has been approved by the FDA for treating acute lymphoid leukemia (ALL), but its therapeutic effect is limited by low catalytic efficiency and L-glutaminase (L-Gln) activity. This study utilized free energy based molecular dynamics calculations to identify residues associated with substrate binding in Bacillus licheniformis L-asparaginase II (BLASNase) with high catalytical activity. After saturation and combination mutagenesis , the mutant LGT (74 L/75G/111 T) with intensively reduced l -glutamine catalytic activity was generated. The l -glutamine/L-asparagine activity (L-Gln/L-Asn) of LGT was only 6.6 % of parent BLASNase, whereas the L-asparagine (L-Asn) activity was preserved >90 %. Furthermore, structural comparison and molecular dynamics calculations indicated that the mutant LGT had reduced binding ability and affinity towards l -glutamine. To evaluate its effect on acute leukemic cells, LGT was supplied in treating MOLT-4 cells. The experimental results demonstrated that LGT was more cytotoxic and promoted apoptosis compared with commercial Escherichia coli ASNase. Overall, our findings firstly provide insights into reducing l -glutamine activity without impacting L-asparagine activity for BLASNase to possess remarkable potential for anti-leukemia therapy.
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2021
·
IF 6.953
Junxin Zhao,Shichang Liu,Yifang Gao,Maomao Ma,Xianghui Yan,Ding Cheng,Dongman Wan,Zheling Zeng,Ping Yu,Deming Gong
Enzymatic degradation has become the most promising approach to degrading organic ester compounds. In this study, Bacillus licheniformis NCU CS-5 was isolated from the spoilage of Cinnamomum camphora seed kernel, and its extracellular lipase was purified, with a specific activity of 192.98 U/mg. The lipase was found to be a trimeric protein as it showed a single band of 27 kDa in SDS-PAGE and 81 kDa in Native-PAGE. It was active in a wide range of temperatures (5–55 °C) and pH values (6.0–9.0), and the optimal temperature and pH value were 40 °C and 8.0, respectively. The enzyme was active in the presence of various organic solvents, metal ions, inhibitors and surfactants. Both crude and purified lipase retained more than 80% activity after 5 h in the presence of commercial detergents, suggesting its great application potential in detergent industry. The highest activity was found to be towards medium- and long-chain fatty acids (C 6 -C 18 ). Peptide mass spectrometric analysis of the purified lipase showed similarity to the lipase family of B. licheniformis . Furthermore, it degraded more than 90% 2,4-D butyl ester to its hydrolysate 2,4-D within 24 h, indicating that the novel lipase may be applied to degrade organic ester pesticides.
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2025
·
IF 6.2
Tianxiang Zhu,Songlin Liu,Yu Gao,Xiongjie Sun,Haiming Hu,Zhigang Zhang,Wenwen Zhang,Lei Xiong,Hongtao Liu
Endoglucanases may not only act on β-1,4-linkages in β-glucan but also target d-glucose in glucomannan, which can expand their utility in the preparation of prebiotics. In this study, a highly efficient endoglucanase (BlGH5) from Bacillus licheniformis BJ2022 was expressed and characterized. BlGH5 exhibited the highest activity at pH 7.0 and 60 °C. It maintained over 80% activity at pH 4.0–7.0 and 30–60 °C. BlGH5 specifically cleaved β-1,4-glycosidic bonds linked to d-glucose. Site-directed mutagenesis results suggested that Arg91, Asn167, Glu168, Trp206, His228, Tyr230, Ser263, and Trp290 are key residues for its binding and catalytic activity. Moreover, BlGH5 displayed high activity against konjac glucomannan (KGM), indicating that BlGH5 could be used to prepare low-molecular-weight konjac glucomannan (KGM-L). Based on the physicochemical properties of KGM and KGM-L, KGM-L was characterized by reduced molecular weight and viscosity. Fecal fermentation experiments demonstrated that KGM and KGM-L could promote the production of short-chain fatty acids (SCFAs). Still, KGM-L was more conducive to the growth of the gut probiotics. In conclusion, we identified an endoglucanase for the degradation of KGM. Results indicated that KGM-L would have superior prebiotic effects. Thus, our study provides a basis for the future development and application of KGM-L as a prebiotic.
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PROCESS BIOCHEMISTRY
·
2024
·
IF 3.7
Yanting Xu,Jing Zhuang,Shuo Wang,Jianying Dai,Zhi-Long Xiu
The aim of this work was to develop an integrated process of 2,3-butanediol production and extraction of active components from Chinese herbs via fermentation, and evaluate their potential as cosmetic ingredients in vitro. A GRAS strain DL2–2 of Bacillus licheniformis was isolated and the fermentation medium was optimized using the ingredients that could be used in cosmetics. Four types of Chinese herbs were added solely or in combination into the fermentation medium to perform 2,3-butanediol production and active components extraction simultaneously. The release of polysaccharides and flavonoids was promoted, and the antioxidant, whitening and antibacterial abilities were improved. The highest antioxidant and antibacterial activity was obtained from the ferment filtrate of Dendrobium devonianum and Gastrodia elata f. glauca , resulting from the highest contents of flavonoids (0.44 g/L) and polyphenols (0.31 g/L), and 42.2 g/L 2,3-butanediol was also produced. The highest polysaccharide concentration was obtained from the ferment filtrate containing Polygonatum kingianum , which exhibited the highest inhibitory effect on tyrosinase (65 %), similar to that of 1 g/L arbutin. Therefore, the filtrate of 2,3-butanediol-Chinese herbal fermentation broth is promising as an ingredient in the production of natural high-value cosmetics.
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BIOCHEMICAL ENGINEERING JOURNAL
·
2025
·
IF 3.8
Wenyu Gao,Guanying Shi,Lihua Zhang,Zhaogai Wang,Wei Zong
γ-glutamyl transpeptidase (GGT, E.C.2.3.2.2) has great applications in the food industry, but there is no commercially available bacterial GGT on the market. In this paper, response surface methodology was used to optimize the yield of Bacillus licheniformis GGT ( Bl GGT). The enzyme was extracted and used to catalyze the synthesis of volatile sulfur compounds (VSCs) in Toona sinensis (TS). The results showed that the optimal conditions for fermentation of GGT production were: lactose 8 g/L, tryptone: soybean meal (1:1) 15 g/L, MgSO 4 ·7 H 2 O 0.5 g/L, K 2 HPO 4 1 g/L, inoculum volume of 5 % v/v, initial pH 8.0, loading volume of 50 mL, and fermentation temperature of 37.0 ℃. Under this condition, the enzyme activity of GGT reached 918.38 ± 0.012 U/L, which was 19 times higher than the activity under the initial medium conditions. In addition, when the GGT was added to the precursor extract from TS, it was noteworthy that VSCs in TS were regained, with the total amount reaching 30.38 μg/mL. The optimization of the fermentation medium to increase the yield of Bl GGT may also provide valuable data for the large-scale production of GGT. The results of this study provide an effective pathway for the synthesis of unique VSCs in TS.
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FOOD CHEMISTRY
·
2021
·
IF 7.514
Junxin Zhao,Maomao Ma,Xianghui Yan,Guohua Zhang,Jiaheng Xia,Guibing Zeng,Wenran Tian,Xianliang Bao,Zheling Zeng,Ping Yu,Deming Gong
A novel lipase from Bacillus licheniformis NCU CS-5 was expressed in different Escherichia coli cells. The recombinant enzyme achieved a high activity (161.74 U/mL) with protein concentration of 0.27 mg/mL under optimal conditions at the large-scale expression of 12 h. The recombinant lipase showed optimal activity at 40 ℃ and pH 10.0, and maintained more than 80% relative activity after 96 h of incubation at pH 9.0–10.0. This typical alkaline lipase was activated under medium temperature conditions (30 and 45 ℃ for 96 h). The lipase exhibited a degree of adaptability in various organic solvents and metal ions, and showed high specificity towards triglycerides with short and medium chain fatty acids. Among different substrates, the lipase showed the strongest binding affinity towards p NPP ( K m = 0.674 mM, V max = 950.196 μM/min). In the experiments of its application in enhancing fatty acids flavor release for low-fat cheeses, the lipase was found to hydrolyze cheeses and mainly increase the contents of butyric acid, hexanoic acid, caprylic acid and decanoic acid. The results from NMR and GC provided the possibility of enhancing fatty acids flavor released from low-fat cheeses by the lipolysis method.
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