High level production of a Bacillus amlyoliquefaciens chitosanase in Pichia pastoris suitable for chitooligosaccharides preparation

Luo, Sa; Qin, Zhen; Chen, Qiming; Fan, Liqiang; Jiang, Lihua; Zhao, Liming

doi:10.1016/j.ijbiomac.2020.02.001

Cited by 37 publications

(51 citation statements)

References 42 publications

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“…AAA19865.1) [ 16 ], Bacillus amyloliquefaciens ECU08 (BaCSN46B, GenBank No. MK257735.1) [ 28 ], Bacillus subtilis 168 (BsCsn, GenBank No. BSU_26890) [ 25 ], Staphylococcus capitis (Csn-CAP, GenBank No.…”

Section: Resultsmentioning

confidence: 99%

“…In comparison with some characterized GH 46 chitosanases ( Table S1 , Supplementary Materials ), as Sn1-CSN most of them exhibited the highest activity below pH 7.0, which were classified as acidic chitosanases, including BaCsn46A from Bacillus amyloliquefaciens YX01 (pH 6.0) [ 14 ], SsCsn46 from Streptomyces sp. N174 (pH 5.5) [ 16 ], BaCSN46B from Bacillus amyloliquefaciens ECU08 (pH 6.5) [ 28 ], BsCsn form Bacillus subtilis 168 (pH 5.0–6.0) [ 25 ], TCH-2 from Bacillus coagulans CK108 (pH 6.5) [ 30 ], BgcsnA from Burkholderia gladioli CHB101(pH 5.6) [ 33 ], Cto1 from Pseudomonas sp. A-01(pH 5.0) [ 35 ], ScCsnA from Streptomyces coelicolor A3(2) (pH 4.1) [ 36 ], CsnA from uncultured bacterium (pH 6.0) [ 37 ], GsCsn46A from Gynuella sunshinyii YC6258(pH 5.5) [ 10 ], BsCsnB from Bacillus sp.…”

Section: Resultsmentioning

confidence: 99%

“…Similar to Sn1-CSN, some GH46 chitosanases were also stable over a wider pH range ( Table S1 , Supplementary Materials ), for example, BaCsn46A(pH 5.0–9.5) [ 14 ], Csn21c (pH 6.0–10.0) [ 17 ], BaCSN46B(pH 3.0–10.5) [ 28 ], GsCsn46A(pH 4.0–9.0) [ 10 ], BsCsn (pH 2.0–9.0) [ 25 ], BsCsnB(pH 3.0–7.5) [ 29 ], and (pH 3.6–9.8)BsCsnQ [ 40 ]. By contrast, some GH46 chitosanases were only stable over a narrow pH range ( Table S1 , Supplementary Materials ), including Csn-BAC(pH 5.5–8.5) [ 3 ], SsCsn46(pH 5.0–6.5) [ 16 ], Csn-CAP (pH 5.0–7.0) [ 29 ], CsnA (pH 4.5–6.5) [ 37 ], Csn-PD (pH 6.0–7.0) [ 38 ] and Cto1 (pH 5.0–8.0) [ 35 ].…”

Section: Resultsmentioning

confidence: 99%

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Expression and Biochemical Characterization of a Novel Marine Chitosanase from Streptomyces niveus Suitable for Preparation of Chitobiose

Chen

Jiao

et al. 2021

Marine Drugs

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It is known that bioactivities of chitooligosaccharide (COS) are closely related to the degree of polymerization (DP); therefore, it is essential to prepare COS with controllable DP, such as chitobiose showing high antioxidant and antihyperlipidemia activities. In this study, BLAST, sequence alignment and phylogenetic analysis of characterized glycoside hydrolase (GH) 46 endo-chitosanases revealed that a chitosanase Sn1-CSN from Streptomyces niveus was different from others. Sn1-CSN was overexpressed in E. coli, purified and characterized in detail. It showed the highest activity at pH 6.0 and exhibited superior stability between pH 4.0 and pH 11.0. Sn1-CSN displayed the highest activity at 50 °C and was fairly stable at ≤45 °C. Its apparent kinetic parameters against chitosan (DDA: degree of deacetylation, >94%) were determined, with Km and kcat values of 1.8 mg/mL and 88.3 s−1, respectively. Cu2+ enhanced the activity of Sn1-CSN by 54.2%, whereas Fe3+ inhibited activity by 15.1%. Hydrolysis products of chitosan (DDA > 94%) by Sn1-CSN were mainly composed of chitobiose (87.3%), whereas partially acetylated chitosan with DDA 69% was mainly converted into partially acetylated COS with DP 2-13. This endo-chitosanase has great potential to be used for the preparation of chitobiose and partially acetylated COS with different DPs.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Expression and Biochemical Characterization of a Novel Marine Chitosanase from Streptomyces niveus Suitable for Preparation of Chitobiose

Chen

Jiao

et al. 2021

Marine Drugs

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“…Several chitosanases have been characterized ( Shinya and Fukamizo 2017 ), they mainly belong to GH 3, GH 5, GH 7, GH 8, GH 46, GH 75, and GH 80 in the Carbohydrate-Active EnZYmes database (CAZy: http://www.cazy.org/ ). Among them, GH family 46 chitosanases are characterized with their excellent catalytic capacity and their potentials to be applied in the production of COS ( Viens et al, 2015 ; Luo et al, 2020 ). Compared with chitosanases, chitinases are much less reported.…”

Section: Introductionmentioning

confidence: 99%

A Novel GH Family 20 β-N-acetylhexosaminidase With Both Chitosanase and Chitinase Activity From Aspergillus oryzae

Zhang

Qin

et al. 2021

Front. Mol. Biosci.

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Aminooligosaccharides possess various biological activities and can exploit wide applications in food, pharmaceutical and cosmetic industries. Commercial aminooligosaccharides are often prepared by the hydrolysis of chitin and chitosan. In this study, a novel GH family 20 β-N-acetylhexosaminidases gene named AoNagase was cloned from Aspergillus oryzae and expressed in Pichia pastoris. The purified AoNagase had maximal activity at pH 5.5 and 65°C. It exhibited good pH stability in the range of pH 6.0–7.5 and at temperatures below 50°C. AoNagase was capable of hydrolyzing not only colloidal chitosan (508.26 U/mg) but also chitin (29.78 U/mg). The kinetic parameters (Km and Vmax) of AoNagase were 1.51 mM, 1106.02 U/mg for chitosan and 0.41 mM, 40.31 U/mg for colloidal chitin. To our knowledge, AoNagase is the first GH family 20 β-N-acetylhexosaminidase capable of hydrolyzing both chitosan and chitin. AoNagase is an endo-type β-N-acetylhexosaminidases and can potentially be used for the manufacturing of aminooligosaccharides.

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“…Chitosanases are divided into seven glycoside hydrolase (GH) families, namely, families 3,5,7,8,46,75 and 80, in accordance with amino acid sequence similarity. Family GH46 chitosanases from Streptomyces (Dubeau et al 2011) and Bacillus (Fukamizo et al 2005) with high catalytic capacity (Luo et al 2020) have been well studied. Crude or puri ed chitosanases are still used for the production of COSs (Sinha et al 2016;Sun et al 2015).…”

Section: Introductionmentioning

confidence: 99%

Evaluation of a Temperature-Induced Chitosanase Bacterial Cell Surface Display System for the Efficient Production of Chitooligosaccharides

Li¹,

Wang²,

Ye³

et al. 2021

Preprint

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Objective To establish a temperature-induced chitosanase bacterial cell surface display system to produce chitooligosaccharides (COSs) efficiently for industrial applications. Results Temperature-inducible chitosanase CSN46A bacterial surface display systems containing one or two copies of ice nucleation protein (InaQ-N) as anchoring motifs were successfully constructed on the basis of Escherichia coli and named as InaQ-N-CSN46A and 2InaQ-N-CSN46A. The specific enzyme activity of 2InaQ-N-CSN46A reached 886.33±0.81 U/g cell dry weight, which was 45.6% higher than that of InaQ-N-CSN46A. However, few proteins were detected in 2InaQ-N-CSN46A hydrolysis system. Therefore, 2InaQ-N-CSN46A had higher hydrolysis efficiency and stability than InaQ-N-CSN46A. GPC revealed that under the optimum enzymatic hydrolysis temperature, the final products were mainly chitobiose and chitotriose. Chitopentaose accumulated (77.62%) when the hydrolysis temperature reached 60 ℃. FTIR and NMR analysis demonstrated that the structures of the two hydrolysis products were consistent with those of COSs.Conclusions In this study, chitosanase was expressed on the surfaces of E. coli by increasing induction temperature, and chitosan was hydrolysed directly without enzyme purification steps. This study provided a novel strategy for industrial COSs production.

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High level production of a Bacillus amlyoliquefaciens chitosanase in Pichia pastoris suitable for chitooligosaccharides preparation

Cited by 37 publications

References 42 publications

Expression and Biochemical Characterization of a Novel Marine Chitosanase from Streptomyces niveus Suitable for Preparation of Chitobiose

Expression and Biochemical Characterization of a Novel Marine Chitosanase from Streptomyces niveus Suitable for Preparation of Chitobiose

A Novel GH Family 20 β-N-acetylhexosaminidase With Both Chitosanase and Chitinase Activity From Aspergillus oryzae

Evaluation of a Temperature-Induced Chitosanase Bacterial Cell Surface Display System for the Efficient Production of Chitooligosaccharides

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