JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2025
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IF 6.2
Jiaojiao Chen,Yiwei Zhang,Mei Zhao,Xinyi Zan,Xiaohong Pan,Cunsheng Zhang,Ziwei Chen,Hossain M. Zabed,Xianghui Qi
Pectate lyase (PL) holds significant potential for applications in various industries. However, the existing PL was unable to adapt to thermo-alkaline industrial environments. In this work, a PL from Caldicellulosiruptor bescii DSM 6725 (hereafter, CbPelD) was studied to disclose its structural and biochemical properties. CbPelD consisted of a putative signal peptide sequence, a short linker region, and a catalytic domain (PelD-II), along with an exoacting β-fructofuranosidase domain (PelD-I). In molecular dynamics simulation, PelD-II demonstrated significantly higher activity and thermal stability compared to other truncated enzymes of CbPelD. The optimal pH and temperature for PelD-II were 10.0 and 60 °C, respectively, with the specific activity increasing by 164–223% in the presence of Ca2+ or Ni2+. The exotype PelD-II exhibited efficient degradation of polygalacturonic acid (PGA) into unsaturated galacturonic acid (uG1), digalacturonic acid (uG2), and trigalacturonic acid (uG3). In the bioscouring assay, PelD-II exhibited a significant increase in the wetted fabric area to 6.072 ± 0.684 cm2, which was about 7.9 times higher than untreated fabric. Scanning electron microscopy further revealed that treatment with PelD-II resulted in a smoother fiber surface, providing direct visual confirmation of the enzyme’s action. These results highlight the potential of PelD-II for applications in the textile industries.
研究领域:
酶学
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生物技术
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纺织工业
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DOI
PubMed
高影响因子
ACS Catalysis
·
2019
·
IF 12.221
Hao Cao,Lichao Sun,Ying Huang,Xin Liu,Dong Yang,Tengfei Liu,Xiaojing Jia,Boting Wen,Tianyi Gu,Fengzhong Wang,Fengjiao Xin
Enzymes are usually characterized by their evolutionarily conserved catalytic domains; however, this work presents the incidental gain-of-function of an enzyme in a loop region by natural evolution of its amino acids. A bifunctional acetyl ester–xyloside hydrolase (CLH10) was heterologously expressed, purified, and characterized. The primary sequence of CLH10 contains the fragments of the conserved sequence of esterase and glycosidase, which distribute in a mixed type. The crystal structure revealed that the primary sequence folded into two independent structural regions to undertake both acetyl esterase and β-1,4-xylanase hydrolase functions. CLH10 is capable of cleaving both the β-1,4-xylosidic bond-linked main chain and the ester bond-linked acetylated side chain of xylan, which renders it valuable because it can degrade acetylated xylan within one enzyme. Significantly, the β-1,4-xylanase activity of CLH10 appears to have been fortuitously obtained because of the variable Asp10 and Glu139 located in its loop region, which suggested that the exposed loop region might act as a potential hot-spot for the design and generation of promising enzyme function in both directed evolution and rational protein design.
研究领域:
生物化学
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酶学
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结构生物学
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分子进化
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DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2017
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IF 3.154
Xin Liu,Tengfei Liu,Yuebin Zhang,Fengjiao Xin,Shuofu Mi,Boting Wen,Tianyi Gu,Xinyuan Shi,Fengzhong Wang,Lichao Sun
Xylanases (EC 3.2.1.8) are a kind of enzymes degrading xylan to xylooligosaccharides (XOS) and have been widely used in a variety of industrial applications. Among them, xylanases from thermophilic microorganisms have distinct advantages in industries that require high temperature conditions. The CoXynA gene, encoding a glycoside hydrolase (GH) family 10 xylanase, was identified from thermophilic Caldicellulosiruptor owensensis and was overexpressed in Escherichia coli. Recombinant CoXynA showed optimal activity at 90 °C with a half-life of about 1 h at 80 °C and exhibited highest activity at pH 7.0. The activity of CoXynA activity was affected by a variety of cations. CoXynA showed distinct substrate specificities for beechwood xylan and birchwood xylan. The crystal structure of CoXynA was solved and a molecular dynamics simulation of CoXynA was performed. The relatively high thermostability of CoXynA was proposed to be due to the increased overall protein rigidity resulting from the reduced length and fluctuation of Loop 7.
研究领域:
生物化学
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微生物学
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结构生物学
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DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2026
·
IF 6.7
Hang Su,Wenjie Han,Hamed I. Hamouda,Huidan Zhang,Ming Lu
β-Xylosidase, crucial for xylan degradation, catalyzes xylose release from xylo-oligosaccharides. However, enzymatic characters of β-xylosidases possess certain limitations, including poor thermal stability, and inhibitors are unclear. This study identified two GH39 family β-xylosidases, CsXyl39A and CsXyl39B, from the thermophilic bacterium Caldicellulosiruptor saccharolyticus. Both enzymes exhibited exceptional thermostability and compatibility with organic solvents, making them promising for industrial applications. CsXyl39A specifically cleaved d-xylopyranosyl groups from substrates such as notoginsenoside R1 and p-nitrophenyl-β-d-xylopyranoside. The catalytic efficiency in organic-solvent/water-miscible (O/Wm) systems was enhanced, and the physical immobilization on macroporous polystyrene strengthened its resistance to organic solvents in high-ratio O/Wm solutions. CsXyl39B demonstrated high specific activity and thermal stability at 70 °C, maintaining a 26 h half-life while producing xylose and glucose, indicating its suitability for lignocellulose biodegradation. Together, these enzymes address key industrial challenges, offering enhanced stability and solvent adaptability for sustainable bioprocessing applications.
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PubMed
高影响因子
BIOPROCESS AND BIOSYSTEMS ENGINEERING
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2023
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IF 3.8
Jia Dong-Xu,Yu Hai,Wang Fan,Jin Li-Qun,Liu Zhi-Qiang,Zheng Yu-Guo
Cellobiose 2-epimerase (CE) is ideally suited to synthesize lactulose from lactose, but the poor thermostability and catalytic efficiency restrict enzymatic application. Herein, a non-characterized CE originating from Caldicellulosiruptor morganii (CmCE) was discovered in the NCBI database. Then, a smart mutation library was constructed based on FoldX ΔΔG calculation and modeling structure analysis, from which a positive mutant D226G located within the α 8 /α 9 loop exhibited longer half-lives at 65–75 °C as well as lower K m and higher k cat / K m values compared with CmCE. Molecular modeling demonstrated that the improvement of D226G was largely attributed to the rigidification of the flexible loop, the compactness of the catalysis pocket and the increment of substrate-binding capability. Finally, the yield of synthesizing lactulose catalyzed by D226G reached 45.5%, higher than the 35.9% achieved with CmCE. The disclosed effect of the flexible loop on enzymatic stability and catalysis provides insight to redesign efficient CEs to biosynthesize lactulose. Graphical abstract
研究领域:
生物技术
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酶学
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分子生物学
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化学工程
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产品关联
DOI
PubMed
高影响因子
Journal of Cleaner Production
·
2021
·
IF 9.297
Guotao Mao,Fangyuan Wang,Jie Wang,Pengyu Chen,Xiaodong Zhang,Hongsen Zhang,Zhimin Wang,Andong Song
Laccases are green biocatalysts that are advantageous in the bioremediation of the triphenylmethane dye malachite green (MG). Here, Cblac from Caldicellulosiruptor bescii was characterized to be a novel DUF152 laccase. Cblac mutant Mut8 was designed with enhanced thermostability, which was demonstrated by the Thermofluor assay, circular dichroism spectrum, and structural analyses. MG bioremediation by Mut8 at high temperatures was performed and examined for its potential to degrade and detoxify the high-temperature MG effluent. Approximately 100% of 100 mg/L of MG was degraded by 40 U/L of Mut8 within 2 h at 75 °C. Mut8 initiated the degradation via oxidative cleavage of the central C–C bond and N-demethylation of MG, and the intermediates were further demethylated, hydroxylated and oxidized. Moreover, the systematic toxicity assessment of the MG degradation products using bacteria, Zea mays , and zebrafish demonstrated that Mut8 thoroughly eliminated MG toxicity. Taken together, Mut8 could effectively decolorize and detoxify MG at high temperatures, showing great promise as an environmental friendly and effective approach to remediate MG effluent immediately released from the dyeing process, thus avoiding the cooling process and lowering the cost simultaneously.
研究领域:
环境生物技术
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生物催化
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生物修复
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酶工程
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产品关联
DOI
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
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2023
·Vol. 107
·期 5
·1751-1764
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IF 5
Hamouda Hamed I.,Fan Yi-Xuan,Abdalla Mohnad,Su Hang,Lu Ming,Li Fu-Li
Abstract Lam16A is a novel GH16 β-1,3–1,4-lichenase isolated from the genus Caldicellulosiruptor which can utilize untreated carbohydrate components of plant cell walls. Its catalytic module has been characterized that the six carbohydrate-binding modules (CBMs) were queued in the C-terminus, but their roles were still unclear. Here, full-length and CBM-truncated mutants of Lam16A were purified and characterized through heterologous expression in Escherichia coli . The profiles of these proteins, including the enzyme activity, degrading efficiency, substrate-binding affinity, and thermostability, were explored. Full-length Lam16A with six CBMs showed excellent thermostability and the highest activity against barley β-glucan and laminarin with optimum pH of 6.5. The CBMs stimulated degrading ability of the catalytic module, especially against β-1,3(4)-glucan-based polysaccharides. The released products from β-1,3–1,4-glucan by Lam16A or its truncated mutants revealed an endo-type glycoside hydrolase. Lam16As exhibited strong binding affinities to the insoluble polysaccharides, especially Lam16A-1CBM. The degradation of yeast cell walls by Lam16A enzyme solution relative to the control reduced the absorbance values at OD 800 by ~ 85% ± 1.2, enabling the release of up to ~ 0.057 ± 0.0039 µg/mL of the cytoplasmic protein into the supernatant, lowering the viability of the cells by ~ 70.3% ± 6.9, thus causing significant damage in the cell wall structure. Taken together, CBMs could influence the substrate specificity, thermal stability, and binding affinity of β-1,3–1,4-glucanase. These results demonstrate the great potential of these enzymes to promote the bioavailability of β-1,3-glucan oligosaccharides for health benefits.
Key points
Carbohydrate-binding modules strongly influenced the enzyme activity and binding affinity, and further impacted glycoside hydrolase activity.
Lam16A enzymes have sufficient ability to hydrolyze β-1,3–1,4-glucan-based polysaccharides.
Lam16As provide a powerful tool to promote the bioavailability of β-1,3-glucan oligosaccharides.
研究领域:
生物化学
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微生物学
·
酶学
·
生物技术
✓ 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）