Characterization of a NaCl-tolerant β-N-acetylglucosaminidase from Sphingobacterium sp. HWLB1

Zhou, Junpei; Song, Zhijun; Zhang, Rui; Limei, Ding; Wu, Qian; Li, Junjun; Tang, Xianghua; Xu, Baojun; Ding, Junmei; Han, Nanyu; Huang, Zunxi

doi:10.1007/s00792-016-0848-4

Cited by 22 publications

(22 citation statements)

References 45 publications

(65 reference statements)

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“…[25]NagA364.424.8 S. thermoviolaceus [83]RmNag321.249.3 Rhizomucor miehei [38]Nag3HWLB1319.419.9 Sphingobacterium sp. [18]NahA38.8- Symbiobacterium thermophilum [47]ExoII30.71.1 V. furnissii [68]NagA3-0.3 Thermotoga maritima [84]CbsA3-0.3 Thermotoga neapolitana [84]--521.5- Enterobacter sp. [85]--67.2- Vibrio alginolyticus [65]--40.5- Talaromyces emersonii [86]--272.0 Streptomyces cerradoensis [40]NagA-5.3245 Pseudomonas fluorescens [3]-0.03- Trichinella spiralis [64]

a These values (μmol min −1 mg −1 ) were determined using p NPGlcNAc as substrate

…”

Section: Resultsmentioning

confidence: 99%

“…[18], the GH 84 GlcNAcase from C. paraputrificum [39] and the GlcNAcase from S. cerradoensis [40]. Ag + and Hg 2+ have irreversible effects on sulfhydryl groups, and Hg 2+ can also oxidize indole rings.…”

Section: Discussionmentioning

confidence: 99%

“…in this study, as well as R. miehei [38], Sphingobacterium sp. [18] and L. casei [41], are strongly inhibited by SDS. Purified rHJ5Nag showed peak activity at pH 6.0 and 45 °C.…”

Section: Discussionmentioning

confidence: 99%

“…An enzyme assay of purified rHJ5Nag towards various substrates was performed spectrophotometrically using the p NP and 3,5-dinitrosalicylic acid (DNS) methods, as described in our previous study [18]. The p NP method was applied to determine the activity of rHJ5Nag towards substrates p NPXyl, p NPGal, p NPGlc, p NPAra, p NPGlcNAc and p NPGalNAc.…”

Section: Methodsmentioning

confidence: 99%

“…HJ5—a new isolate from the saline soil. The purified recombinant HJ5Nag (rHJ5Nag), which was expressed in E. coli , showed better tolerance to salts and GlcNAc and higher activity than most GlcNAcases reported in the literature, including the GH 3 GlcNAcase we previously reported [18]. Potential molecular adaptations for salt tolerance and the high activity of the enzyme were presumed.…”

Section: Introductionmentioning

confidence: 99%

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Distinctive molecular and biochemical characteristics of a glycoside hydrolase family 20 β-N-acetylglucosaminidase and salt tolerance

et al. 2017

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BackgroundEnzymatic degradation of chitin has attracted substantial attention because chitin is an abundant renewable natural resource, second only to lignocellulose, and because of the promising applications of N-acetylglucosamine in the bioethanol, food and pharmaceutical industries. However, the low activity and poor tolerance to salts and N-acetylglucosamine of most reported β-N-acetylglucosaminidases limit their applications. Mining for novel enzymes from new microorganisms is one way to address this problem.ResultsA glycoside hydrolase family 20 (GH 20) β-N-acetylglucosaminidase (GlcNAcase) was identified from Microbacterium sp. HJ5 harboured in the saline soil of an abandoned salt mine and was expressed in Escherichia coli. The purified recombinant enzyme showed specific activities of 1773.1 ± 1.1 and 481.4 ± 2.3 μmol min−1 mg−1 towards p-nitrophenyl β-N-acetylglucosaminide and N,N'-diacetyl chitobiose, respectively, a V max of 3097 ± 124 μmol min−1 mg−1 towards p-nitrophenyl β-N-acetylglucosaminide and a K i of 14.59 mM for N-acetylglucosamine inhibition. Most metal ions and chemical reagents at final concentrations of 1.0 and 10.0 mM or 0.5 and 1.0% (v/v) had little or no effect (retaining 84.5 − 131.5% activity) on the enzyme activity. The enzyme can retain more than 53.6% activity and good stability in 3.0–20.0% (w/v) NaCl. Compared with most GlcNAcases, the activity of the enzyme is considerably higher and the tolerance to salts and N-acetylglucosamine is much better. Furthermore, the enzyme had higher proportions of aspartic acid, glutamic acid, alanine, glycine, random coils and negatively charged surfaces but lower proportions of cysteine, lysine, α-helices and positively charged surfaces than its homologs. These molecular characteristics were hypothesised as potential factors in the adaptation for salt tolerance and high activity of the GH 20 GlcNAcase.ConclusionsBiochemical characterization revealed that the GlcNAcase had novel salt–GlcNAc tolerance and high activity. These characteristics suggest that the enzyme has versatile potential in biotechnological applications, such as bioconversion of chitin waste and the processing of marine materials and saline foods. Molecular characterization provided an understanding of the molecular–function relationships for the salt tolerance and high activity of the GH 20 GlcNAcase.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-017-0358-1) contains supplementary material, which is available to authorized users.

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a These values (μmol min −1 mg −1 ) were determined using p NPGlcNAc as substrate

…”

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…in this study, as well as R. miehei [38], Sphingobacterium sp. [18] and L. casei [41], are strongly inhibited by SDS. Purified rHJ5Nag showed peak activity at pH 6.0 and 45 °C.…”

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Distinctive molecular and biochemical characteristics of a glycoside hydrolase family 20 β-N-acetylglucosaminidase and salt tolerance

et al. 2017

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Biotransformation of Chitinous Waste into Value-Added Products

Kumar

Vivekanand

Pareek

2020

Environmental Microbiology and Biotechnology

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Enzymatic properties of β-N-acetylglucosaminidases

Zhang

Zhou

Song

et al. 2017

Appl Microbiol Biotechnol

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β-N-Acetylglucosaminidases (GlcNAcases) hydrolyse N-acetylglucosamine-containing oligosaccharides and proteins. These enzymes produce N-acetylglucosamine (GlcNAc) and have a wide range of promising applications in the food, energy, and pharmaceutical industries, such as synergistic degradation of chitin with endo-chitinases and using GlcNAc to produce sialic acid, bioethanol, single-cell proteins, and pharmaceutical therapeutics. GlcNAcases also play an important role in the dynamic balance of cellular O-linked GlcNAc levels, catabolism of ganglioside storage in Tay-Sachs disease, and bacterial cell wall recycling and flagellar assembly. In view of these important biological functions and the wide range of industrial applications of GlcNAcases, this review aims to provide a better understanding of various advances for these enzymes. It focuses on enzymatic properties of GlcNAcases, including substrate specificity, catalytic activity, pH optimum, temperature optimum, thermostability, the effects of various metal ions and organic reagents, and transglycosylation.

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Characterization of a NaCl-tolerant β-N-acetylglucosaminidase from Sphingobacterium sp. HWLB1

Cited by 22 publications

References 45 publications

Distinctive molecular and biochemical characteristics of a glycoside hydrolase family 20 β-N-acetylglucosaminidase and salt tolerance

Distinctive molecular and biochemical characteristics of a glycoside hydrolase family 20 β-N-acetylglucosaminidase and salt tolerance

Biotransformation of Chitinous Waste into Value-Added Products

Enzymatic properties of β-N-acetylglucosaminidases

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