Improving the thermostability of a GH11 xylanase by directed evolution and rational design guided by B-factor analysis.

Abstract:

:Operational stability under high temperature is required for enzyme application in industrial processes. Error-prone PCR and B-factor analysis were employed to enhance the thermostability of a xylanase from GH family 11 in this study. Based on the top 10 mutants screened from the random mutation libraries, mutant Xyn371 was derived from the optimal mutant Xyn370 by integrating the beneficial residues identified in the other 9 screened mutants. Subsequently, a best-saturation mutant Xyn372 originated from Xyn371 was selected with a 60-min half-life at 70 °C (0.5-min half-life for the wild-type enzyme). According to the site-saturated mutagenesis of 10 residues with higher B-factors in Xyn372, mutants Xyn375 and Xyn376 were screened; their half-lives at 70 °C were 410 and 360 min, respectively. The substituted residues located in the "palm" region of the N-terminus and the newly generated hydrogen bonds in the mutants might contribute to improved thermostability. The significantly improved thermostability of mutants will pave the way for applications in different industrial areas.

journal_name

Enzyme Microb Technol

authors

Xing H,Zou G,Liu C,Chai S,Yan X,Li X,Liu R,Yang Y,Zhou Z

doi

10.1016/j.enzmictec.2020.109720

subject

Has Abstract

pub_date

2021-02-01 00:00:00

pages

109720

eissn

0141-0229

issn

1879-0909

pii

S0141-0229(20)30213-1

journal_volume

143

pub_type

杂志文章
  • Improvement of catalytical properties of two invertases highly tolerant to sucrose after expression in Pichia pastoris. Effect of glycosylation on enzyme properties.

    abstract::Zymomonas mobilis genes encoding INVA and INVB were expressed in Pichia pastoris, under the control of the strong AOX1 promoter, and the recombinant enzymes were named INVAAOX1 and INVBAOX1. The expression levels of INVAAOX1 (1660 U/mg) and INVBAOX1 (1993 U/mg) in P. pastoris were 9- and 7-fold higher than those obser...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2015.11.008

    authors: Pérez de los Santos AI,Cayetano-Cruz M,Gutiérrez-Antón M,Santiago-Hernández A,Plascencia-Espinosa M,Farrés A,Hidalgo-Lara ME

    更新日期:2016-02-01 00:00:00

  • Utilization of distillery wastewater for hydrogen production in one-stage and two-stage processes involving photofermentation.

    abstract::In this study, distillery wastewater was treated by dark fermentation or photofermentation alone, and by sequential dark and photofermentation processes using anaerobic saccharolytic consortium and purple nonsulfur bacteria. Combination of dark and photofermentation resulted in the maximal H2 yield of 17.6L/L of disti...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2017.11.009

    authors: Laurinavichene T,Tekucheva D,Laurinavichius K,Tsygankov A

    更新日期:2018-03-01 00:00:00

  • A thermophilic enzymatic cocktail for galactomannans degradation.

    abstract::The full utilization of hemicellulose sugars (pentose and exose) present in lignocellulosic material, is required for an efficient bio-based fuels and chemicals production. Two recombinant thermophilic enzymes, an endo-1,4-β-mannanase from Dictyoglomus turgidum (DturCelB) and an α-galactosidase from Thermus thermophil...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2017.12.008

    authors: Aulitto M,Fusco FA,Fiorentino G,Bartolucci S,Contursi P,Limauro D

    更新日期:2018-04-01 00:00:00

  • Efficient production of gluten hydrolase Kuma030 in E. coli by hot acid treatment without chromatography.

    abstract::Kumamolisin from Alicyclobacillus sendaiensis strain NTAP-1 is a serine protease with collagenase activity. After molecular engineering, a kumamolisin mutant, named Kuma030, was obtained with high proteolytic activity against gluten, which might cause celiac disease. Kuma030 exhibited its potential application in indu...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2019.05.013

    authors: Liu H,Fan X,Song H,Hu X,Zhang G,Yu C,Yi L

    更新日期:2019-10-01 00:00:00

  • Antibacterial activity of emulsions containing unsaturated fatty acid ergosterol esters synthesized by lipase-mediated transesterification.

    abstract::Ergosterol, like cholesterol, has many beneficial physiological activities, and because it has no adverse clinical problem with cholesterol, it is a substance that can be used as a biomaterial in the food and cosmetics industry. However, ergosterol has low oil solubility and is easily crystallized, which is problemati...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2020.109581

    authors: Park SH,Kim HK

    更新日期:2020-09-01 00:00:00

  • Glucosylation of the flavonoid, astragalin by Leuconostoc mesenteroides B-512FMCM dextransucrase acceptor reactions and characterization of the products.

    abstract::Astragalin (kaempferol-3-O-β-D-glucopyranoside, Ast) glucosides were synthesized by the acceptor reaction of a dextransucrase produced by Leuconostoc mesenteroides B-512FMCM with astragalin and sucrose. Each glucoside was purified and their structures were assigned as kaempferol-3-O-β-D-glucopyranosyl-(1→3)-O-α-D-gluc...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2011.09.007

    authors: Kim GE,Kang HK,Seo ES,Jung SH,Park JS,Kim DH,Kim DW,Ahn SA,Sunwoo C,Kim D

    更新日期:2012-01-05 00:00:00

  • Sucrose hydrolysis by thermostable immobilized inulinases from aspergillus ficuum.

    abstract::The possibility of using thermostable inulinases from Aspergillus ficuum in place of invertase for sucrose hydrolysis was explored. The commercial inulinases preparation was immobilized onto porous glass beads by covalent coupling using activation by a silane reagent and glutaraldehyde before adding the enzyme. The im...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/s0141-0229(00)00342-2

    authors: Ettalibi M,Baratti JC

    更新日期:2001-05-07 00:00:00

  • Overexpression of NAD kinases improves the L-isoleucine biosynthesis in Corynebacterium glutamicum ssp. lactofermentum.

    abstract::NADPH is the key cofactor in L-isoleucine (Ile) biosynthetic pathway. To increase the Ile biosynthesis in Corynebacterium glutamicum ssp. lactofermentum JHI3-156, NADPH supply needs to be enhanced. Here NAD kinase, the key enzyme for the de novo biosynthesis of NADP(+) and NADPH, were cloned and expressed in JHI3-156,...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2012.04.003

    authors: Shi F,Huan X,Wang X,Ning J

    更新日期:2012-07-15 00:00:00

  • The boosting effect of recombinant hemicellulases on the enzymatic hydrolysis of steam-treated sugarcane bagasse.

    abstract::To increase the efficiency of enzyme cocktails in deconstructing cellulose and hemicelluloses present in the plant cell wall, a combination of enzymes with complementary activities is required. Xylan is the main hemicellulose component of energy crops and for its complete hydrolysis a system consisting of several enzy...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2019.109447

    authors: Cintra LC,da Costa IC,de Oliveira ICM,Fernandes AG,Faria SP,Jesuíno RSA,Ravanal MC,Eyzaguirre J,Ramos LP,de Faria FP,Ulhoa CJ

    更新日期:2020-02-01 00:00:00

  • Effect of protein load on stability of immobilized enzymes.

    abstract::Different lipases have been immobilized on octyl agarose beads at 1mg/g and at maximum loading, via physical interfacial activation versus the octyl layer on the support. The stability of the preparations was analyzed. Most biocatalysts had the expected result: the apparent stability increased using the highly loaded ...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2016.12.002

    authors: Fernandez-Lopez L,Pedrero SG,Lopez-Carrobles N,Gorines BC,Virgen-Ortíz JJ,Fernandez-Lafuente R

    更新日期:2017-03-01 00:00:00

  • Optimization of the growth conditions of Sf21 insect cells for high-density perfusion culture in stirred-tank bioreactors.

    abstract::Spodoptera frugiperda insect cells (IPLB-Sf21-AE) (Sf21), infected with baculovirus expression vectors during their exponential growth phase, are commonly used to produce a variety of heterologous recombinant proteins. In the present study the culture conditions of these insect cells were studied to establish high-den...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/0141-0229(94)90022-1

    authors: Deutschmann SM,Jäger V

    更新日期:1994-06-01 00:00:00

  • A new pH indicator dye-based method for rapid and efficient screening of l-asparaginase producing microorganisms.

    abstract::l-asparaginase is a pharmaceutically and industrially important enzyme as it has potential to treat different cancers and inhibit acrylamide formation in fried and baked food products. In the present study, an attempt was made to screen for new and novel l-asparaginase producers using a widely applied phenol red and b...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2017.08.004

    authors: Mihooliya KN,Nandal J,Swami L,Verma H,Chopra L,Sahoo DK

    更新日期:2017-12-01 00:00:00

  • Direct and selective enzymatic synthesis of trehalose unsaturated fatty acid diesters and evaluation of foaming and emulsifying properties.

    abstract::Trehalose diesters are Gemini-type surfactants that might have better surface activity than conventional surfactants. A one-step method for the preparation of trehalose unsaturated fatty acid diesters has been successfully developed. The yield of trehalose diester of different unsaturated fatty acids was between 78 % ...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2020.109516

    authors: Ji S,Jia C,Cao D,Li S,Zhang X

    更新日期:2020-05-01 00:00:00

  • Cellulase-lignin interactions-the role of carbohydrate-binding module and pH in non-productive binding.

    abstract::Non-productive cellulase adsorption onto lignin is a major inhibitory mechanism preventing enzymatic hydrolysis of lignocellulosic feedstocks. Therefore, understanding of enzyme-lignin interactions is essential for the development of enzyme mixtures and processes for lignocellulose hydrolysis. We have studied cellulas...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2013.07.003

    authors: Rahikainen JL,Evans JD,Mikander S,Kalliola A,Puranen T,Tamminen T,Marjamaa K,Kruus K

    更新日期:2013-10-10 00:00:00

  • Peptide synthesis by proteases in organic solvents: medium effect on substrate specificity.

    abstract::The substrate specificities of alpha-chymotrypsin and subtilisins for peptide synthesis in hydrophilic organic solvents were investigated. Chymotrypsin exhibited high specificity to aromatic amino acids as acyl donors, while subtilisin Carlsberg and subtilisin BPN' were specific to aromatic and neutral aliphatic amino...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/0141-0229(92)90102-t

    authors: Nagashima T,Watanabe A,Kise H

    更新日期:1992-10-01 00:00:00

  • Eco-friendly intracellular biosynthesis of CdS quantum dots without changing Escherichia coli's antibiotic resistance.

    abstract::In the paper, a green and efficient biosynthetical technique was reported for preparing cadmium sulfide (CdS) quantum dots, in which Escherichia coli (E. coli) was chosen as a biomatrix. Fluorescence emission spectra and fluorescent microscopic photographs revealed that as-produced CdS quantum dots had an optimum fluo...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2016.09.017

    authors: Yan ZY,Du QQ,Qian J,Wan DY,Wu SM

    更新日期:2017-01-01 00:00:00

  • Enzyme reaction engineering: synthesis of antibiotics catalysed by stabilized penicillin G acylase in the presence of organic cosolvents.

    abstract::By using very active and very stable penicillin G acylase (PGA)--agarose derivatives we have studied the industrial design of equilibrium-controlled synthesis of lactamic antibiotics. In the presence of high concentrations of organic cosolvents we have carried out the direct enzymatic condensation of phenylacetic acid...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/0141-0229(91)90106-k

    authors: Fernandez-Lafuente R,Rosell CM,Guisán JM

    更新日期:1991-11-01 00:00:00

  • The use of oxygen uptake rate measurements to control the supply of toxic substrate: toluene hydroxylation by Pseudomonas putida UV4.

    abstract::During toluene hydroxylation, catalyzed by Pseudomonas putida UV4 one molecule of oxygen is added to the aromatic ring to produce the dihydroxylated (non-aromatic) ring structure, toluene cis-glycol. Toluene, which is toxic to the cells at aqueous phase concentration above ( approximately 2.4 mmol), is fed to the reac...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/s0141-0229(00)00289-1

    authors: Carragher JM,McClean WS,Woodley JM,Hack CJ

    更新日期:2001-02-01 00:00:00

  • The application of foaming for the recovery of Surfactin from B. subtilis ATCC 21332 cultures.

    abstract::Foaming, a proficient method for the recovery of surface active solutes from dilute solutions, was successfully applied for the concentration of the lipopeptide biosurfactant Surfactin from B. subtilis ATCC 21332 cell culture broths. Foaming was only partially successful in concentrating Surfactin when applied as a se...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/s0141-0229(00)00327-6

    authors: Davis DA,Lynch HC,Varley J

    更新日期:2001-03-08 00:00:00

  • Optimization of sodium dedecyl sulfate (SDS) addition coupled with adenosine triphosphate (ATP) regeneration for glutathione overproduction in high density cultivation of Candida utilis.

    abstract::In the previous study, glutathione production was elevated by adding precursor amino acids and adenosine triphosphate (ATP) in high cell density cultivation of Candida utilis. Furthermore, in the present research, glutathione production was further improved by optimizing sodium dedecyl sulfate (SDS) addition coupled w...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2010.02.007

    authors: Liang G,Mo Y,Du G

    更新日期:2010-05-05 00:00:00

  • Comments on "Bioprocessing in space"

    abstract::An analysis developed by Westgate et al. for the digestible energy of edible and inedible biomass, including hydrolysis and fermentation, is reexamined with state-of-the-art values for the harvest index of hydroponic crops. ...

    journal_title:Enzyme and microbial technology

    pub_type: 评论,杂志文章

    doi:10.1016/0141-0229(93)90104-a

    authors: Volk T

    更新日期:1993-10-01 00:00:00

  • Reaction strategies for the enzymatic synthesis of neopentyl glycol diheptanoate.

    abstract::This work describes for the first time the green synthesis of neopentyl glycol diheptanoate in a solvent-free medium via an enzymatic pathway. The process has been carried out in an open-air reactor in order to ease water removal through evaporation and shift the chemical equilibrium towards product formation. The inh...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2019.109400

    authors: Serrano-Arnaldos M,García-Martínez JJ,Ortega-Requena S,Bastida J,Máximo F,Montiel MC

    更新日期:2020-01-01 00:00:00

  • Increasing the refolding efficiency in vitro by site-directed mutagenesis of Cys383 in rat procarboxypeptidase B.

    abstract::This study examines a novel method to reduce the probability of disulfide mismatches during the refolding process by the replacement of cysteines within a protein. Specifically, Cys383 of recombinant rat procarboxypeptidase B was replaced by other amino acids to increase the refolding efficiency in vitro. Mutants C383...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2011.04.018

    authors: Li S,Zhang L,Wu Q,Xin A,Zhao J,Fan L

    更新日期:2011-07-10 00:00:00

  • Efficient enzymatic synthesis of α-keto acids by redesigned substrate-binding pocket of the l-amino acid deaminase (PmiLAAD).

    abstract::In our previous study, we produced α-keto acids by using an L-amino acid deaminase PmiLAAD (wide-type) from Proteus mirabilis, however, the catalytic efficiency was low due to its low substrate affinity. In this study, protein engineering of PmiLAAD was performed to improve the α-keto acid production. PmiLAAD was engi...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2019.109393

    authors: Wu L,Guo X,Wu G,Liu P,Liu Z

    更新日期:2020-01-01 00:00:00

  • A new strategy for strain improvement of Aurantiochytrium sp. based on heavy-ions mutagenesis and synergistic effects of cold stress and inhibitors of enoyl-ACP reductase.

    abstract::Developing a strain with high docosahexaenoic acid (DHA) yield and stable fermenting-performance is an imperative way to improve DHA production using Aurantiochytrium sp., a microorganism with two fatty acid synthesis pathways: polyketide synthase (PKS) pathway and Type I fatty acid synthase (FAS) pathway. This study ...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2016.08.019

    authors: Cheng YR,Sun ZJ,Cui GZ,Song X,Cui Q

    更新日期:2016-11-01 00:00:00

  • Use of raw glycerol to produce oil rich in polyunsaturated fatty acids by a thraustochytrid.

    abstract::Glucose is the typical carbon source for producing microbial polyunsaturated fatty acids (PUFA) with single cell microorganisms such as thraustochytrids. We assessed the use of a fish oil derived glycerol by-product (raw glycerol), produced by a fish oil processing plant, as a carbon source to produce single cell oil ...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2010.11.008

    authors: Scott SD,Armenta RE,Berryman KT,Norman AW

    更新日期:2011-03-07 00:00:00

  • An efficient purification process for sweet potato beta-amylase by affinity precipitation with alginate.

    abstract::beta-amylases are used in production of maltose syrup. It is shown that sweet potato beta-amylase can be purified by affinity precipitation with alginate with 80% activity yield and 44 fold purification. SDS-PAGE of the purified protein showed a single band and a subunit weight of 50 kDa. Preliminary data with soybean...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/s0141-0229(01)00338-6

    authors: Teotia S,Khare SK,Gupta MN

    更新日期:2001-06-07 00:00:00

  • Purification and functional characterization of the first stilbene glucoside-specific β-glucosidase isolated from Lactobacillus kimchi.

    abstract::This study aimed to develop viable enzymes for bioconversion of resveratrol-glucoside into resveratrol. Out of 13 bacterial strains tested, Lactobacillus kimchi JB301 could completely convert polydatin into resveratrol. The purified enzyme had an optimum temperature of 30-40°C and optimum pH of pH 5.0 against polydati...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2014.09.001

    authors: Ko JA,Park JY,Kwon HJ,Ryu YB,Jeong HJ,Park SJ,Kim CY,Oh HM,Park CS,Lim YH,Kim D,Rho MC,Lee WS,Kim YM

    更新日期:2014-12-01 00:00:00

  • The development and application of a new affinity partitioning system for enzyme isolation and purification.

    abstract::A reactive water-soluble polymer was synthesized by copolymerizing N-isopropylacrylamide and glycidyl acrylate. The reactive polymer could react with the amino groups of enzymes/proteins or other ligands to form an affinity polymer. As a model, the reactive polymer was allowed to react with paraaminobenzamidine, a str...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/0141-0229(90)90005-b

    authors: Nguyen AL,Luong JH

    更新日期:1990-09-01 00:00:00

  • Amphiphilic hollow carbonaceous microsphere-encapsulated enzymes: Facile immobilization and robust biocatalytic properties.

    abstract::This study reports a general strategy for the encapsulation of various enzymes in amphiphilic hollow carbonaceous microspheres (CMs). We found that enzymes could be spontaneously encapsulated in the interior cavity of the CMs via hydrophobic interactions. Due to strong hydrophobic interactions and robust confinement, ...

    journal_title:Enzyme and microbial technology

    pub_type: 杂志文章

    doi:10.1016/j.enzmictec.2015.06.015

    authors: Liu L,Sun Y,He L,Jiang L,Yang S

    更新日期:2015-11-01 00:00:00