The Draft Genome Sequence and Analysis of an Efficiently Chitinolytic Bacterium Chitinibacter sp. Strain GC72

Zhang, Alei; Mo, Xiaofang; Wei, Guoguang; Zhou, Ning; Yang, Suping; Chen, Jie; Wang, Yingying; Chen, Kequan; Ouyang, Pingkai

doi:10.1007/s00284-020-02215-9

Cited by 7 publications

(8 citation statements)

References 22 publications

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“…(57.5 kDa), V. harveyi 650 (73 kDa) [26]and S. thermoviolaceus (60 kDa) [31]. However, the M w of NAGaseA is similar with the previously reported GH20 NAGase from C. meiyuanensis with a molecular mass of 92,571Da [32]. The speci c activity of recombinant NAGaseA exhibited a 1.39-fold increase from 270.17 U/mg to 373.29 U/mg with a protein recovery of 78.6% yield after puri cation (Table S1).…”

Section: Expression Of Nagasea Gene and Puri Cation Of Recombinant Nagaseasupporting

confidence: 83%

“…F-3 (60℃) [34], and P. hydrolytica S66 (50°C) [35]. As for the thermostability pro le, the activity dropped rapidly after incubation at temperatures above 40ºC, suggesting the poor thermostability of NAGaseA, which were similar with that of GH20 GlcNAGases from C. meiyuanensis [32], Microbacterium sp. HJ5 [27] and Aeromonas sp.…”

Section: Expression Of Nagasea Gene and Puri Cation Of Recombinant Nagaseamentioning

confidence: 82%

“…These results indicated that NAGaseA capable of producing higher N-acetyl COSs ((GlcNAc) 3 −(GlcNAc) 6 ) from (GlcNAc) 2 −(GlcNAc) 5 , which exhibited trans glycosylation activity. Our previous study also reported that CmNAGase from Chitinolyticbacter meiyuanensis SYBC-H1 could produce higher N-acetyl COSs (GlcNAc) 3 −(GlcNAc) 7 from (GlcNAc) 2 −(GlcNAc) 6 , respectively [32]. However, unlike CmNAGase, no new peak presumed as (GlcNAc) 7 generated when using (GlcNAc) 6 as the substrate, which could be attributed to the lower reverse hydrolysis activity of NAGaseA.…”

Section: Effects Of Metal Ions On Activity Of Recombinant Nagaseamentioning

confidence: 88%

“…The temperature and pH pro les of recombinant NAGaseA were investigated in Fig 3 . As shown in Fig. 3(A), the recombinant NAGaseA displayed the optimal temperature at 40℃, which was consisted with NAGase from C. Meiyuanensis SYBC-H1 (40℃) [32], but different from NAGases from S. marcescens (52°C) [33], Microbacterium sp. HJ5 (45℃) [27], Streptomyces sp.…”

Section: Expression Of Nagasea Gene and Puri Cation Of Recombinant Nagaseamentioning

confidence: 99%

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Characterization of New Multifunctional GH20 β-N-Acetylglucosaminidase From Chitinibacter sp. GC72 and Its Application in Converting Chitin Into N-Acetyl Glucosamine

Chen

Zhou

Chen

et al. 2022

Preprint

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Background N-acetyl glucosamine (GlcNAc), possesses many specific bioactivities and has been widely used in biomedical, food, and chemical industries. Alternatively, enzymatic hydrolysis of chitin into GlcNAc using chitinolytic enzymes was shown to be a more attractive approach in recent years, because of the green process and the excellent bioactivity of product. Therefore, it is of economic and environmental value to realize the efficient production of GlcNAc from abundant chitin resources. Results In this study, a gene encoding β-N-acetylglucosaminidase, designated NAGaseA, was cloned from Chitinibacter sp. GC72 and subsequently functional expressed in Escherichia coli BL21 (DE3). NAGaseA contains a glycoside hydrolase family 20 catalytic domain that shows low identity with the corresponding domain of the well-characterized NAGases. The recombinant NAGaseA had a molecular mass of 92 kDa. Biochemical characterization of the purified NAGaseA revealed that the optimal reaction condition was at 40°C and pH 6.5, and exhibited great pH stability in the range of pH 6.5–9.5. The Vmax, Km, Kcat, and Kcat/Km of NAGaseA toward pNP-GlcNAc were 3333.33 µmol min−1 L−1, 39.99 µM, 4667.07 s−1, and 116.71 mL µmol −1 s−1, respectively. Analysis of the hydrolysis products of N-acetyl chitinoligosaccharides (N-Acetyl COSs) indicated that NAGaseA capable of converting N-Acetyl COSs((GlcNAc)2–(GlcNAc)6) into GlcNAc with hydrolysis ability order: (GlcNAc)2 > (GlcNAc)3 > (GlcNAc)4 > (GlcNAc)5 > (GlcNAc)6. Moreover, NAGaseA could generate (GlcNAc)3–(GlcNAc)6 from (GlcNAc)2–(GlcNAc)5, respectively. These results showed that NAGaseA is a multi-functional NAGase with transglycosylation activity. In addition, significantly synergistic action was observed between NAGaseA and other sources of chitinases during hydrolysis of colloid chitin. Finally, 0.759 g/L, 0.481 g/L and 0.986 g/L of GlcNAc with purity of 96% were obtained using different chitinase combinations, resulting in 1.6-2.69 fold increase, respectively. Conclusions The hydrolytic properties and good environmental adaptions indicate that NAGaseA has great potential in the bioconversion of chitin waste and behaved as an excellent candidate in GlcNAc producing.

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Section: Expression Of Nagasea Gene and Puri Cation Of Recombinant Nagaseasupporting

confidence: 83%

Section: Expression Of Nagasea Gene and Puri Cation Of Recombinant Nagaseamentioning

confidence: 82%

Section: Effects Of Metal Ions On Activity Of Recombinant Nagaseamentioning

confidence: 88%

Section: Expression Of Nagasea Gene and Puri Cation Of Recombinant Nagaseamentioning

confidence: 99%

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Characterization of New Multifunctional GH20 β-N-Acetylglucosaminidase From Chitinibacter sp. GC72 and Its Application in Converting Chitin Into N-Acetyl Glucosamine

Chen

Zhou

Chen

et al. 2022

Preprint

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“…GC72 isolated from pond mud in Nanjing were capable of hydrolyzing chitin into GlcNAc as the sole product ( Gao et al, 2015 ). Moreover, only one gene encoding NAGase, named NAGaseA, was found in strain GC72 via complete genome sequencing and analysis ( Zhang et al, 2020a ). In this study, the NAGaseA gene was cloned from the genome of strain GC72 and heterologously expressed in Escherichia coli (BL21).…”

Section: Introductionmentioning

confidence: 99%

Characterization of a New Multifunctional GH20 β-N-Acetylglucosaminidase From Chitinibacter sp. GC72 and Its Application in Converting Chitin Into N-Acetyl Glucosamine

et al. 2022

Self Cite

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In this study, a gene encoding β-N-acetylglucosaminidase, designated NAGaseA, was cloned from Chitinibacter sp. GC72 and subsequently functional expressed in Escherichia coli BL21 (DE3). NAGaseA contains a glycoside hydrolase family 20 catalytic domain that shows low identity with the corresponding domain of the well-characterized NAGases. The recombinant NAGaseA had a molecular mass of 92 kDa. Biochemical characterization of the purified NAGaseA revealed that the optimal reaction condition was at 40°C and pH 6.5, and exhibited great pH stability in the range of pH 6.5–9.5. The Vmax, Km, kcat, and kcat/Km of NAGaseA toward p-nitrophenyl-N-acetyl glucosaminide (pNP-GlcNAc) were 3333.33 μmol min–1 l–1, 39.99 μmol l–1, 4667.07 s–1, and 116.71 ml μmol–1 s–1, respectively. Analysis of the hydrolysis products of N-acetyl chitin oligosaccharides (N-Acetyl COSs) indicated that NAGaseA was capable of converting N-acetyl COSs ((GlcNAc)2–(GlcNAc)6) into GlcNAc with hydrolysis ability order: (GlcNAc)2 > (GlcNAc)3 > (GlcNAc)4 > (GlcNAc)5 > (GlcNAc)6. Moreover, NAGaseA could generate (GlcNAc)3–(GlcNAc)6 from (GlcNAc)2–(GlcNAc)5, respectively. These results showed that NAGaseA is a multifunctional NAGase with transglycosylation activity. In addition, significantly synergistic action was observed between NAGaseA and other sources of chitinases during hydrolysis of colloid chitin. Finally, 0.759, 0.481, and 0.986 g/l of GlcNAc with a purity of 96% were obtained using three different chitinase combinations, which were 1.61-, 2.36-, and 2.69-fold that of the GlcNAc production using the single chitinase. This observation indicated that NAGaseA could be a potential candidate enzyme in commercial GlcNAc production.

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Chemo-enzymatic protocol converts chitin into a nitrogen-containing furan derivative, 3-acetamido-5-acetylfuran

Chen

Wang

et al. 2021

Molecular Catalysis

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The Draft Genome Sequence and Analysis of an Efficiently Chitinolytic Bacterium Chitinibacter sp. Strain GC72

Cited by 7 publications

References 22 publications

Characterization of New Multifunctional GH20 β-N-Acetylglucosaminidase From Chitinibacter sp. GC72 and Its Application in Converting Chitin Into N-Acetyl Glucosamine

Characterization of New Multifunctional GH20 β-N-Acetylglucosaminidase From Chitinibacter sp. GC72 and Its Application in Converting Chitin Into N-Acetyl Glucosamine

Characterization of a New Multifunctional GH20 β-N-Acetylglucosaminidase From Chitinibacter sp. GC72 and Its Application in Converting Chitin Into N-Acetyl Glucosamine

Chemo-enzymatic protocol converts chitin into a nitrogen-containing furan derivative, 3-acetamido-5-acetylfuran

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