APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2020
·Vol. 104
·期 11
·4927-4943
·
IF 3.53
Deng Pengjun,Meng Chunyu,Wu Yan,Xu Jie,Tang Xiaoqing,Zhang Xuecheng,Xiao Yazhong,Wang Xiaotang,Fang Zemin,Fang Wei
A novel β -glucosidase, BglD1 with high β -galactosidase and transglycosidation activities, was screened and cloned from the deep-sea bacterium Bacillus sp. D1. BglD1 exhibited the maximal β -glucosidase and β -galactosidase activities at 55–60 °C and pH 5.5–6.0. The enzyme maintained approximately 50% of its original activity at 35 °C and pH 6.0 after 120-h incubation. When applied to synthesize galacto-oligosaccharides (GOS), BglD1 generated 118.3 g/L GOS (33.8% (w/w)) from 350 g/L lactose, with trisaccharide Gal- β (1 → 3)-Lac and disaccharide Gal- β (1 → 4)-Gal as the main components. Furthermore, BglD1 could hydrolyze lactose in milk and produce GOS simultaneously. Using milk as the substrate, BglD1 hydrolyzed 88.5% lactose and produced 3.3 g/L GOS after incubation at 30 °C for 1 h. To improve the transglycosidation activity, a mutant BglD1:E224T was generated based on the semi-rational design. The GOS yield of BglD1:E224T was 11.5% higher than that of BglD1 when using lactose solution as the substrate. Thus, BglD1 and the mutant could be used as beneficial alternatives of the existing β -galactosidases for the production of GOS.
研究领域:
微生物学
·
酶工程
·
食品科学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF FOOD BIOCHEMISTRY
·
2019
·Vol. 43
·期 11
·e13027
·
IF 1.358
Mengnan Hong,Jing Li,Yuwen Chen
This work aim to study the tolerance and aroma-related enzymes activities of the non- Saccharomyces yeasts in Vidal blanc icewine from the Huanren region of China. The strains were identified by sequencing internal transcribed spacer (ITS) region and the 26S rDNA D1/D2 domain genes and all representative non- Saccharomyces yeasts were belonged to genera Metschnikowia , Hanseniaspora , Torulaspora, Candida , and Debaryomyces . A total of 28 strains were carried out for tolerance experiments and results suggested that most of them could tolerate 500 g/L glucose, 4% ethanol, 20 g/L tartaric acid, and 350 mg/L SO 2 . Finally, a total of 17 strains with better tolerance were carried out for the β-glucosidase, β-xylosidase, and pectinase activities experiments. The results showed that Candida railenensis HC08 and the strains of Hanseniaspora genus have satisfactory multi-enzyme activities, which can be used to design mixed fermentation to produce characteristic icewine. Practical applications Non- Saccharomyces yeasts produces a series of hydrolytic enzymes that are thought to have a significant contribution to the aroma complexity of wines, however, are poorly explored in icewine. In this work, most of the non- Saccharomyces yeasts screened from Chinese icewine can adapt well to the high-sugar and high-acid environment of icewine, and can secrete hydrolase. The application of these strains in mixed fermentation could provide a prospect for the production of characteristic icewine.
研究领域:
酿酒学
·
微生物学
·
食品科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2017
·Vol. 183
·期 4
·1488-1502
·
IF 1.751
Wang Liming,Chen Yan
Cycloastragenol (CA), an exclusive telomerase activator, was derived from the Astragali Radix which is widely distributed in Turkey. Until now, there is no report to produce CA with effective and environment-friendly methods. Biotransformation is considered to be a promising technology. Thus, the present study was aimed to establish a biotransformation technology that could efficiently produce CA. In this paper, a microorganism, LG-502, was used to successfully transform astragaloside IV (ASI) to CA by analysis of thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC). The phylogenetic analysis of the 16S rRNA indicated that this strain belongs to Bacillus sp. Three metabolites were separated during the fermentation and characterized to be cyclogaleginoside B, CA, and 20 R , 24 S -epoxy-6 α , 16 β , 25-trihydroxy-9, 19-cycloartan-3-one based on NMR and MS spectroscopic analyses. The conversion rate of ASI and yield rate of CA were achieved as high as 89 and 84%, respectively, under optimized conditions. Enzymatic analysis showed that the glycosidases were mainly located inside the bacterial body, and the activities of glucosidases were much higher than the xylosidases under the experimental conditions. This study provides a feasible, effective, and eco-friendly way to prepare CA from ASI, which might greatly contribute to the applications of ASI.
研究领域:
药学
·
生物技术
·
微生物学
·
有机化学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF FOOD PROCESSING AND PRESERVATION
·
2021
·Vol. 45
·期 7
·e15604
·
IF 2.19
Cheng Peng,Rui Li,Hui Ni,Li Jun Li,Qing Biao Li
The naringinase, a desirable candidate for citrus juice debittering, consists of α-L-rhamnosidase and β-D-glucosidase. This study aimed to elucidate the effects of α-L-rhamnosidase, β-D-glucosidase, and their combination on naringin, polyphenols, antioxidant activity, and aroma quality. The α-L-rhamnosidase and the two enzymes combination reduced the naringin concentration from 356.33 to 10.52 and 4.17 mg/L, respectively. The α-L-rhamnosidase, β-D-glucosidase, and their combination significantly increased the antioxidant activity. The unpleasant cooking note was significantly weakened and the floral note was increased after three treatments. The combinational treatment increased the sweet note greatly. The substrate competition between α-L-rhamnosidase and β-D-glucosidase may help to explain the inconsistencies of the polyphenols, antioxidant activity, and aroma quality after single enzyme or combined enzymes treatments. The results indicated that taste, antioxidant activity, and aromatic quality were well improved by the combined treatment than either α-L-rhamnosidase or β-D-glucosidase treatment, providing an attractive reference for debittering orange juice. Practical applications The work showed that the combined treatment of α-L-rhamnosidase and β-D-glucosidase was better to remove the bitter taste of citrus juice than the solo treatment with α-L-rhamnosidase or β-D-glucosidase in regarding with taste, antioxidant activity, and aromatic quality. It helps to better the quality of citrus juice with enzymatic treatment.
研究领域:
食品科学
·
酶学
·
饮料技术
✓ 已验证
产品关联
DOI
高影响因子
MOLECULES
·
2015
·Vol. 20
·期 4
·6601-6610
·
IF 2.416
Jie-Tao Pan,Bang-Wei Yu,Yong-Qin Yin,Jie-Hong Li,Li Wang,Li-Bing Guo,Zhi-Bin Shen
Six pentasaccharide resin glycosides fromIpomoea cairica, including four new acylated pentasaccharide resin glycosides, namely cairicoside I–IV (1–4) and the two known compounds cairicoside A (5) and cairicoside C (6), were isolated from the aerial parts ofIpomoea cairica. Their structures were established by a combination of spectroscopic, including two dimensional (2D) NMR and chemical methods. The core of the six compounds was simonic acid A, and they were esterfied the same sites, just differing in the substituent groups. The lactonization site of the aglycone was bonded to the second saccharide moiety at C-2 in1–4, and at C-3 in5–6. Compounds1and5,4and6were two pairs of isomers. The absolute configuration of the aglycone in1–6which was (11S)-hydroxyhexadecanoic acid (jalapinolic acid) was established by Mosher’s method. Compounds1–4have been evaluated for inhibitory activity against α-glucosidase, which all showed inhibitory activities.Keywords:Ipomoea cairica;resin glycoside;simonic acid A;α-glucosidase
研究领域:
植物化学
·
药理学
✓ 已验证
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Jingjing Zhang,Jing Huang,Yufan Yang,Daxin Zhao,Cong Wang,Peng Zhong,Jianhua Jia,Weifan Dang,Qiwei Lu,Chengyu Zhang,Xiaohui Yan
Icaritin (ICT), a compound with diverse biological activities derived from Epimedium folium, is typically present in low concentrations in EFs. However, the abundant glycosyl-modified ICT compounds facilitate its transformation into ICT. Current biocatalytic production faces challenges, including low conversion rates and limited enzyme activity. This study developed a one-pot enzymatic cascade strategy for directly biotransform crude extracts of Epimedium folium (EEF) to produce ICT. The feasibility of catalyzing different ICT-related compounds in EEF was validated through molecular docking and substrate reactions. The selected glycosidase exhibited simultaneous activities as a glucosidase, xylosidase, and α-1,6-rhamnosidase, with the rhamnosidase showing outer-rhamnosidic activity and weak glucosidase activity. By using EFs as the substrate and employing whole-cells ( Escherichia coli ) containing LacS and BtRha proteins for synergistic catalysis, icariin can be efficiently synthesized within 6 h, achieving a conversion rate of 100 %. The enzymatic cascade for ICT production from crude extracts was elucidated by analyzing catalytic intermediates via HPLC. Compared to strategies using single or traditional multi-enzyme applications, this method shows advantages of ease to operation, high efficiency, and large production yield performance. This method has the potential to become an eco-friendly catalytic strategy for the large-scale production of icaritin.
研究领域:
生物技术
·
生物化学
·
生药学
·
酶学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Berry Research
·
2019
·Vol. 9
·期 1
·109-123
·
IF 2.379
Zhang Jiaojiao,Xiao Jianbo,Giampieri Francesca,Forbes-Hernandez Tamara Y.,Gasparrini Massimiliano,Afrin Sadia,Cianciosi Danila,Reboredo-Rodriguez Patricia,Battino Maurizio,Zheng Xiaodong
Anthocyanins are plant pigments widely present in fruits and vegetables that exert health benefits as part of human diet. They exist in not glycosylated forms (anthocyanidins) and glycosylated forms. Anthocyanins have been studied for their ability to inhibit polysaccharide metabolism enzymes (such as α-glucosidase) and, therefore, attenuate hyperglycemia and modulate type II diabetes. The aim of the present work was to compare the inhibitory activity of anthocyanins with that of acarbose, a common α-glucosidase inhibitor, by evaluating IC50 and Km, to compare the inhibitory capabilities among different compounds in an in vitro study. Our results showed that most of the anthocyanins studied presented lower IC50 values than acarbose; only Pn-3-glc presented IC50 value close to acarbose. Moreover, there was no difference in Km between complete competitive and complete non-competitive types of inhibition (T-test, p > 0.05). In conclusion, the anthocyanins and their degradation products work together on attenuating abilities of this sugar metabolism key enzyme, throughout competitive and non-competitive modes.
研究领域:
营养学
·
药理学
✓ 已验证
产品关联
DOI
高影响因子
Food Bioscience
·
2024
·
IF 4.8
Fuhao Tang,Baoyao Wei,Chao Qin,Li Huang,Ning Xia,Jianwen Teng
Passion fruit peel contained various phenolic compounds with α-glucosidase and α-amylase inhibitory activities in vitro . However, the bound state and glycosylated forms hindered their active expression. In this article, β-glucosidase-producing Lactobacilli were inoculated into fermented peels for targeted metabolism. The results showed that the release of bound polyphenols and the increase of free polyphenols were significantly correlated with β-glucosidase activity. The strain G1 with high enzyme activity degraded 48.20% of the bound polyphenols after 24 h. The ability of extracts to inhibit α-glucosidase and α-amylase activities was significantly enhanced and closely related to the content of free polyphenols (r = -0.96 and -0.88, P < 0.001). Computer- assisted screening identified 38 phenolic compounds as potential dual inhibitors of α-glucosidase and α-amylase. Among them, 22 α-glucosidase inhibitors and 24 α-amylase inhibitors were identified for the first time. Various inhibitors, including phenolic acids, flavones, and flavonols, increased in content after fermentation. The total content of 14 glycosylated polyphenols decreased to 28.38% at 48 h, which was opposite to the aglycone. Molecular docking indicated that glycosylated polyphenols bind to β-glucosidase through hydrogen bonds and van der Waals forces. Different glycosylations at the C-3 position of quercetin bound at same sites to β-glucosidase. This study contributes to the theory of enhancing the functional activity of natural products through targeted fermentation transformation.
研究领域:
食品科学
·
生物化学
·
微生物学
·
药理学
✓ 已验证
DOI
高影响因子
Microbial Cell Factories
·
2023
·Vol. 22
·期 1
·1-11
·
IF 6.4
Lin Yu,Chen Wen-wen,Ding Bo,Guo Man,Liang Meng,Pang Hao,Wei Yu-tuo,Huang Ri-bo,Du Li-qin
Background Icaritin is an aglycone of flavonoid glycosides from Herba Epimedii. It has good performance in the treatment of hepatocellular carcinoma in clinical trials. However, the natural icaritin content of Herba Epimedii is very low. At present, the icaritin is mainly prepared from flavonoid glycosides by α-L-rhamnosidases and β-glucosidases in two-step catalysis process. However, one-pot icaritin production required reported enzymes to be immobilized or bifunctional enzymes to hydrolyze substrate with long reaction time, which caused complicated operations and high costs. To improve the production efficiency and reduce costs, we explored α-L-rhamnosidase SPRHA2 and β-glucosidase PBGL to directly hydrolyze icariin to icaritin in one-pot, and developed the whole-cell catalytic method for efficient icaritin production.Results The SPRHA2 and PBGL were expressed in Escherichia coli, respectively. One-pot production of icaritin was achieved by co-catalysis of SPRHA2 and PBGL. Moreover, whole-cell catalysis was developed for icariin hydrolysis. The mixture of SPRHA2 cells and PBGL cells transformed 200 g/L icariin into 103.69 g/L icaritin (yield 95.23%) in 4 h in whole-cell catalysis under the optimized reaction conditions. In order to further increase the production efficiency and simplify operations, we also constructed recombinant E. coli strains that co-expressed SPRHA2 and PBGL. Crude icariin extracts were also efficiently hydrolyzed by the whole-cell catalytic system.Conclusion sCompared to previous reports on icaritin production, in this study, whole-cell catalysis showed higher production efficiency of icaritin. This study provides promising approach for industrial production of icaritin in the future.
研究领域:
生物技术
·
药学
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Natural Products and Bioprospecting
·
2024
·Vol. 14
·期 1
·1-16
·
IF 4.8
Yang Ya-Ya,Jing Shun-Li,Shao Jia-Li,Chen Ji-Xuan,Zhang Wei-Feng,Wan Si-Yuan,Shen Yu-Ping,Yang Huan,Yu Wei
Functionalized mesoporous materials have become a promising carrier for enzyme immobilization. In this study, Santa Barbara Amorphous 15 (SBA-15) was modified by N-aminoethyl- γ -aminopropyl trimethoxy (R). R-SBA-15 was employed to purify and immobilize recombinant β -glucosidase from Terrabacter ginsenosidimutans (BgpA) in one step for the first time. Optimum pH of the constructed R-SBA-15@BgpA were 7.0, and it has 20 ℃ higher optimal temperature than free enzyme. Relative activity of R-SBA-15@BgpA still retained > 70% at 42 ℃ after 8-h incubation. The investigation on organic reagent resistance revealed that the immobilized enzyme can maintain strong stability in 15% DMSO. In leaching test and evaluation of storage stability, only trace amount of protein was detected in buffer of the immobilized enzyme after storage at 4 ℃ for 33 days, and the immobilized BgpA still maintained > 50% relative activity. It also demonstrated good reusability, with 76.1% relative activity remaining after fourteen successive enzymatic hydrolyses of epimedin A to sagittatoside A. The newly proposed strategy is an effective approach for the purification and immobilization of BgpA concurrently. In addition, R-SBA-15@BgpA was demonstrated to have high efficiency and stability in this application, suggesting its great feasibility and potential to produce bioactive compounds such as secondary glycosides or aglycones from natural products. Graphical Abstract
研究领域:
生物化学
·
酶工程
·
材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD HYDROCOLLOIDS
·
2020
·
IF 7.053
Mengting Ni,Xing Hu,Deming Gong,Guowen Zhang
Vitexin, a C-glycosylated flavonoid , is abundant in numerous foods . In this study, vitexin was determined as an uncompetitive α-glucosidase inhibitor and its binding site was different from the active site. It possessed a remarkably higher inhibitory activity than acarbose with its IC 50 value of 52.80 ± 1.65 μM. Interestingly, the combination of acarbose with vitexin showed a strong synergistic inhibition effect. The stable complex between vitexin and α-glucosidase was mainly formed by hydrogen bonds and hydrophobic forces. The presence of vitexin induced a looser conformation of α-glucosidase and decreased its surface hydrophobicity. Computational docking and chemical modification of α-glucosidase found that residues Glu, Asn, Lys, and Thr were essential for the binding course and their importance order was Glu, Asn > Lys > Thr. Molecular dynamics simulation results revealed the molecular mechanism of the interaction between vitexin and α-glucosidase and verified the above experimental results. This work proposed rewarding findings in functional research of vitexin as a potential α-glucosidase inhibitor.
研究领域:
食品科学
·
生物化学
·
药理学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF MEDICINAL CHEMISTRY
·
2024
·
IF 6.8
Xin Xiang,Chuning Dong,Lianbo Zhou,Jun Liu,Zachary M. Rabinowitz,Yuzhao Zhang,Honghui Guo,Feng He,Xingdou Chen,Yunhua Wang,Lina Cui,Xiaowei Ma
Real-time detection of cellular senescence remains a clinical challenge. Here, we aimed to develop a positron emission tomography (PET) imaging probe targeting senescence-associated β-galactosidase (SA-β-Gal), the most widely used biomarker of cellular senescence, and investigate its performance for real-time in vivo quantitative detection of cellular senescence. A stable PET imaging agent [68Ga]Ga-BGal was obtained with a high labeling yield (90.0 ± 4.3%) and a radiochemical purity (>95%). [68Ga]Ga-BGal displayed high sensitivity and specificity for β-Gal both in vitro and in vivo. The reaction and uptake of the probe correlated with the β-Gal concentration and reaction time. In PET imaging, high β-Gal-expressing CT26.CL25 tumors and doxorubicin-treated HeLa tumors showed high signals from [68Ga]Ga-BGal, while a low signal was observed in CT26.WT and untreated HeLa tumors. In summary, we showcased successful PET imaging of senescence in preclinical models using probe [68Ga]Ga-BGal. This finding holds the potential for translating senescence imaging into clinical applications.
研究领域:
医学影像学
·
分子生物学
·
肿瘤学
·
放射学
·
药学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）