Marine Drugs
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2021
·Vol. 19
·期 11
·596
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IF 5.118
Lin-Lin Zhang,Xiao-Hua Jiang,Xin-Feng Xiao,Wen-Xiu Zhang,Yi-Qian Shi,Zhi-Peng Wang,Hai-Xiang Zhou
(1) Background: Chitooligosaccharides (COS) have numerous applications due to their excellent properties. Chitosan hydrolysis using chitosanases has been proposed as an advisable method for COS preparation. Although many chitosanases from various sources have been identified, the cold-adapted ones with high stability are still rather rare but required. (2) Methods: A novel chitosanase named CsnY from marine bacteriumRenibacteriumsp. Y82 was expressed inEscherichia coli, following sequence analysis. Then, the characterizations of recombinant CsnY purified through Ni–NTA affinity chromatography were conducted, including effects of pH and temperature, effects of metal ions and chemicals, and final product analysis. (3) Results: The GH46 family chitosanase CsnY possessed promising thermostability at broad temperature range (0–50 °C), and with optimal activity at 40 °C and pH 6.0, especially showing relatively high activity (over 80% of its maximum activity) at low temperatures (20–30 °C), which demonstrated the cold-adapted property. Common metal ions or chemicals had no obvious effect on CsnY except Mn2+and Co2+. Finally, CsnY was determined to be an endo-type chitosanase generating chitodisaccharides and -trisaccharides as main products, whose total concentration reached 56.74 mM within 2 h against 2% (w/v) initial chitosan substrate. (4) Conclusions: The results suggest the cold-adapted CsnY with favorable stability has desirable potential for the industrial production of COS.Keywords:chitosanase;cold adaptation;chitooligosaccharide;Renibacteriumsp.
研究领域:
生物化学
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生物技术
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海洋生物学
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DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2018
·
IF 4.946
Zhen Qin,Qiming Chen,Si Lin,Sa Luo,Yongjun Qiu,Liming Zhao
Chitooligosaccharide is widely used as a functional food additive and a valuable pharmacological agent. The transformation of chitinous biomass into valuable bioactive chitooligosaccharides is one of the most exciting applications of chitosanase. A novel glycoside hydrolase (GH) family 46 chitosanase (GsCsn46A) from rhizobacterium Gynuella sunshinyii was cloned and heterologously expressed in Escherichia coli . GsCsn46A showed maximal activity at pH 5.5 and 30 °C. GsCsn46A featured remarkable cold-adapted property, which controllably hydrolyzed chitosan to three types of chitooligosaccharides at the mild reaction condition (reaction condition: pH 5.5 at 30 °C; method for stopping the reaction: 50 °C for 30 min). The yields of three types of chitooligosaccharides products (degree of polymerization (DP): 2–7, 2–5 and 2–3) were 70.9%, 87.1% and 94.6% respectively. This novel cold-adapted chitosanase provides a cleaner production process for the controllable preparation of chitooligosaccharides with the specific DP.
研究领域:
生物化学
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酶工程
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食品科学
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DOI
PubMed
高影响因子
JOURNAL OF SURFACTANTS AND DETERGENTS
·
2024
·
IF 1.6
Qinzhou Cai,Huifang Zhang,Bingshan Zhao,Lei Ren,Yonghua Wang,Fanghua Wang
Cold-adapted, alkali-stable lipases with surfactant tolerance are highly sought after in the detergent industry. In this study, we recombinantly expressed and characterized a novel lipase from Marinobacter nanhaiticus (MNL). The purified MNL exhibited high stability within a temperature range of 10–30°C and pH values of 8–10, demonstrating optimal activity at 20°C and pH 8. MNL has a preference for substrates with medium chains with the best activity being 3768.6 U/mg using p -nitrophenyl decanoate as a substrate. Remarkably, MNL showed enhanced enzymatic activity in the presence of ionic surfactants and displayed notable resilience when exposed to proteases. Washing performance analysis further revealed MNL's high proficiency in removing oil-based stains from fabrics. Collectively, these results suggest that MNL holds significant promise as a valuable component in laundry detergent formulations.
研究领域:
生物化学
·
工业微生物学
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DOI
高影响因子
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY
·
2020
·
IF 4.187
Yanbo Wang,Feifei Wang,Xiaoshuang Zhang,Congnan Cen,Linglin Fu
Shewanella baltica is the specific spoilage microorganism of Pseudosciaena crocea during low-temperature storage. Exploring the correlation between cold adaptability and spoilage potential may provide a new perspective for prolonging shelf life of aquatic products. In the present study, we investigated the synthesis pathway of unsaturated fatty acid (UFA) responsible for regulating cold adaptability in Shewanella baltica and its effect on spoilage potential. FabR and FadR, as key regulators of membrane unsaturated fatty acids synthesis pathway, were identified in S. baltica . FabR was significantly down-regulated at 4 °C compared to at 30 °C, yet FadR displayed the opposite results. By overexpressing fabR and fadR genes at 4 °C, we found that FabR and FadR had negative and positive effects on UFA content and membrane fluidity as well as spoilage potential, respectively. These data indicated that FabR and FadR functioned collectively to increase the membrane fluidity for better cold adaptability at low temperature, resulting in the maintenance of spoilage potential of S. baltica .
研究领域:
微生物学
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食品科学
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分子生物学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2023
·
IF 8.2
Jianrong Chen,Dengfeng Yang,Yunkai Zhang,Liyan Yang,Qingyan Wang,Mingguo Jiang,Lixia Pan
In order to better utilize chitinolytic enzymes to produce high-value N -acetyl-D-glucosamine (GlcNAc) from chitinous waste, there is an urgent need to explore bi-functional chitinases with exceptional properties of temperature, pH and metal tolerance. In this study, we cloned and characterized a novel bi-functional cold-adaptive chitinase called Ca Chi18A from a newly isolated strain, Chitinilyticum aquatile CSC-1, in Bama longevity village of Guangxi Province, China. The activity of Ca Chi18A at 50 °C was 4.07 U/mg. However, it exhibited significant catalytic activity even at 5 °C. Its truncated variant Ca Chi18A_ΔChBDs, containing only catalytic domain, demonstrated significant activity levels, exceeding 40 %, over a temperature range of 5–60 °C and a pH range of 3 to 10. It was noteworthy that it displayed tolerance towards most metal ions at a final concentration of 0.1 mM, including Fe 3+ and Cu 2+ ions, retaining 122.52 ± 0.17 % and 116.42 ± 1.52 % activity, respectively. Additionally, it exhibited favorable tolerance towards organic solvents with the exception of formic acid. Interestedly, Ca Chi18A and Ca Chi18A_ΔChBDs had a low K m value towards colloidal chitin (CC), 0.94 mg mL −1 and 2.13 mg mL −1 , respectively. Both enzymes exhibited chitobiosidase and N -acetyl-D-glucosaminidase activities, producing GlcNAc as the primary product when hydrolyzing CC. The high activities across a broader temperature and pH range, strong environmental adaptability, and hydrolytic properties of Ca Chi18A_ΔChBDs suggested that it could be a promising candidate for GlcNAc production.
研究领域:
生物化学
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酶工程
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环境科学
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PubMed
高影响因子
MOLECULES
·
2019
·Vol. 24
·期 21
·3915
·
IF 3.06
Yue Yang,Zhou Zheng,Yifei Xiao,Jiaojiao Zhang,Yu Zhou,Xiao Li,Shangyong Li,Huiqing Yu
Chitosanase plays an important role in the production of chitooligosaccharides (CHOS), which possess various biological activities. Herein, a glycoside hydrolase (GH) family 46 chitosanase-encoding gene,csnB, was cloned from marine bacteriumBacillussp. BY01 and heterologously expressed inEscherichia coli. The recombinant chitosanase, CsnB, was optimally active at 35 °C and pH 5.0. It was also revealed to be a cold-adapted enzyme, maintaining 39.5% and 40.4% of its maximum activity at 0 and 10 °C, respectively. Meanwhile, CsnB showed wide pH-stability within the range of pH 3.0 to 7.0. Then, an improved reaction condition was built to enhance its thermostability with a final glycerol volume concentration of 20%. Moreover, CsnB was determined to be an endo-type chitosanase, yielding chitosan disaccharides and trisaccharides as the main products. Overall, CsnB provides a new choice for enzymatic CHOS production.Keywords:chitosanase;Bacillussp. BY01;cold-adaption
研究领域:
酶学
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海洋生物学
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生物技术
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DOI
PubMed
高影响因子
JOURNAL OF BIOTECHNOLOGY
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2020
·
IF 3.503
Shi-Yu Zhang,Ze-Wang Guo,Xiao-Ling Wu,Xiao-Yang Ou,Min-Hua Zong,Wen-Yong Lou
Cold-adapted pullulanase with high catalytic activity and stability is of special interest for its wide application in cold starch hydrolysis, but few pullulanases displaying excellent characteristics at ambient temperature and acidic pH have hitherto been reported. Here, a novel pullulanase from Bacillus methanolicus PB1 was successfully expressed in Escherichia coli BL21 (DE3) and determined to be a cold-adapted type I pullulanase (PulPB1) with maximum activity at 50 °C and pH 5.5. The recombinant PulPB1 showed great stability, its half-life at 50 °C was 137 h. PulPB1 can efficiently hydrolyze pullulan and amylopectin, with activities of 292 and 184 U/mg at 50 °C and pH 5.5, respectively. Moreover, the N-terminal domain of PulPB1 was found to significantly affect the enzymatic performance. Following truncation of the N-terminal domain, the activity towards pullulan decreased markedly from 292 to 141 U/mg and the half-life at 50 °C decreased from 137 to 10 h. Compared to the hydrolysis system with amyloglucosidase alone, the catalytic efficiency showed a 2.4-fold increase on combining PulPB1 with amyloglucosidase for amylopectin hydrolysis at 40 °C. This demonstrates that PulPB1 is promising for development as a superior candidate for cold amylopectin hydrolysis.
研究领域:
生物化学
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微生物学
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酶学
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生物技术
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DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2024
·
IF 5.7
Liyan Yang,Mingbo Qu,Zhou Wang,Shiyong Huang,Qingyan Wang,Maochun Wei,Fei Li,Dengfeng Yang,Lixia Pan
GH19 (glycoside hydrolase 19) chitinases play crucial roles in the enzymatic conversion of chitin and biocontrol of phytopathogenic fungi. Herein, a novel multifunctional chitinase of GH19 (CaChi19A), which contains three chitin-binding domains (ChBDs), was successfully cloned from Chitinilyticum aquatile CSC-1 and heterologously expressed in Escherichia coli. We also generated truncated mutants of CaChi19A_ΔI, CaChi19A_ΔIΔII, and CaChi19A_CatD consisting of two ChBDs and a catalytic domain, one ChBD and a catalytic domain, and only a catalytic domain, respectively. CaChi19A, CaChi19A_ΔI, CaChi19A_ΔIΔII, and CaChi19A_CatD exhibited cold adaptation, as their relative enzyme activities at 5 °C were 40.7, 51.6, 66.2, and 82.6%, respectively. Compared with CaChi19A and other variants, CaChi19A_ΔIΔII demonstrated a higher level of stability below 50 °C and retained relatively high activity over a wide pH range of 5–12. Analysis of the hydrolysis products revealed that CaChi19A and CaChi19A_ΔIΔII exhibit exoacting, endoacting, and N-acetyl-β-d-glucosaminidase activities toward colloidal chitin. Furthermore, CaChi19A and CaChi19A_ΔIΔII exhibited inhibitory effects on the hyphal growth of Fusarium oxysporum, Fusarium redolens, Fusarium fujikuroi, Fusarium solani, and Coniothyrium diplodiella, thereby illustrating effective biocontrol activity. These results indicated that CaChi19A and CaChi19A_ΔIΔII show advantages in some applications where low temperatures were demanded in industries as well as the biocontrol of fungal diseases in agriculture.
研究领域:
微生物学
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酶学
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植物病理学
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生物技术
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产品关联
DOI
PubMed
高影响因子
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