Theoretical and experimental studies on the conformational changes of organic solvent-stable protease from Bacillus sphaericus DS11 in methanol/water mixtures

Gu, Zhanghui; Lai, Jiangli; Hang, Jiahao; Zhang, Chunguang; Wang, Shujun; Jiao, Yuliang; Liu, Shu; Fang, Yiwu

doi:10.1016/j.ijbiomac.2019.01.196

Cited by 31 publications

(24 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…conducted molecular dynamics simulations to investigate how methanol molecules were able to enter into the active site of an organic solvent-stable protease, leading to microenvironmental changes around the aromatic amino acids. Their results suggest that high concentrations of methanol lead to the exposure of hydrophobic groups to the solvent, leading to decrease in enzyme activity and stability 54 . Since aromatic and hydrophobic amino acids are very sensitive to organic solvents, Loo et al .…”

Section: Discussionmentioning

confidence: 99%

“…These relevant studies indicated the microenvironment changes caused by organic solvents may be pivotal for the regulation of enzyme activity. The molecular dynamics simulation/determination and electrospray-ionization (ESI) mass spectrometry could be used to more effectively reveal the mechanism underlying the phenomenon described in this work, which could be a good topic for future studies 54,57 .…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Enhancement of laccase activity by pre-incubation with organic solvents

Lin

Lee

et al. 2019

Sci Rep

View full text Add to dashboard Cite

Laccases that are tolerant to organic solvents are powerful bio-catalysts with broad applications in biotechnology. Most of these uses must be accomplished at high concentration of organic solvents, during which proteins undergo unfolding, thereby losing enzyme activity. Here we show that organic-solvent pre-incubation provides effective and reversible 1.5- to 4.0-fold enhancement of enzyme activity of fungal laccases. Several organic solvents, including acetone, methanol, ethanol, DMSO, and DMF had an enhancement effect among all laccases studied. The enhancement was not substrate-specific and could be observed by using both phenolic and non-phenolic substrates. Laccase preincubated with organic solvents was sensitive to high temperature but remained stable at 25 °C, for an advantage for long-term storage. The acetone-pre-incubated 3-D structure of DLac, a high-efficiency fungal laccase, was determined and confirmed that the DLac protein structure remains intact and stable at a high concentration of organic solvent. Moreover, the turnover rates of fungal laccases were improved after organic-solvent pre-incubation, with DLac showing the highest enhancement among the fungal laccases examined. Our investigation sheds light on improving fungal laccase usage under extreme conditions and extends opportunities for bioremediation, decolorization, and organic synthesis.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Enhancement of laccase activity by pre-incubation with organic solvents

Lin

Lee

et al. 2019

Sci Rep

View full text Add to dashboard Cite

show abstract

“…Recombinant E. coli KRX/pCon2(3) harboring the organic solvent tolerant elastase strain K gene was obtained from the Enzyme and Microbial Technology (EMTech) Research Centre, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia. Overexpression of elastase strain K was conducted according to Rahman et al [11]. The bacterial culture was then sonicated thrice with 30% duty cycle and 8 control output settings to harvest the enzyme.…”

Section: Bacterial and Enzyme Sourcementioning

confidence: 99%

“…Most of these enzymes are secreted by bacterial strains isolated from organic-solvent-contaminated sites such as Pseudomonas sp., Bacillus sp., Rhizopus oryzae, Halobacterium sp., and Natrialba magadi. There are also reports on thermostable enzyme that is able to tolerate the presence of 25% (v/v) organic solvents and is said to have an enhanced catalytic activity [10][11][12][13]. The effect of organic solvent on enzymes may accentuate the activity of the enzyme in this situation; the organic solvent molecules interact with the enzyme at different polarities and organic solvent concentrations which improve the conformation of the enzyme.…”

Section: Introductionmentioning

confidence: 99%

Biochemical and Structural Characterization of Cross-Linked Enzyme Aggregates (CLEAs) of Organic Solvent Tolerant Protease

et al. 2020

View full text Add to dashboard Cite

Cross-linked enzyme aggregates (CLEAs) is an immobilization technique that can be used to customize enzymes under an optimized condition. Structural analysis on any enzyme treated with a CLEA remains elusive and has been less explored. In the present work, a method for preparing an organic solvent tolerant protease using a CLEA is disclosed and optimized for better biochemical properties, followed by an analysis of the structure of this CLEA-treated protease. The said organic solvent tolerant protease is a metalloprotease known as elastase strain K in which activity of the metalloprotease is measured by a biochemical interaction with azocasein. Results showed that when a glutaraldehyde of 0.02% (v/v) was used under a 2 h treatment, the amount of recovered activity in CLEA-elastase was highest. The recovered activity of CLEA-elastase and CLEA-elastase-SB (which was a CLEA co-aggregated with starch and bovine serum albumin (BSA)) were at an approximate 60% and 80%, respectively. The CLEA immobilization of elastase strain K allowed the stability of the enzyme to be enhanced at high temperature and at a broader pH. Both CLEA-elastase and CLEA-elastase-SB end-products were able to maintain up to 67% enzyme activity at 60 °C and exhibiting an enhanced stability within pH 5–9 with up to 90% recovering activity. By implementing a CLEA on the organic solvent tolerant protease, the characteristics of the organic solvent tolerant were preserved and enhanced with the presence of 25% (v/v) acetonitrile, ethanol, and benzene at 165%, 173%, and 153% relative activity. Structural analysis through SEM and dynamic light scattering (DLS) showed that CLEA-elastase had a random aggregate morphology with an average diameter of 1497 nm.

show abstract

“…The RMSD represents the relative positional shift of the protease structure at various times and the overall fluctuation level of protein conformation, based on its initial construction 39 . The stability of L. fermentum R6 protease under conditions of pH 7 amd 40 °C and pH 8 and 70 °C can be reflected by RMSD analysis.…”

Section: Resultsmentioning

confidence: 99%

Effects of temperature and pH on the structure of a metalloprotease from Lactobacillus fermentumR6 isolated from Harbin dry sausages and molecular docking between protease and meat protein

et al. 2021

View full text Add to dashboard Cite

BACKGROUND Microbial protease can interact with meat protein in fermented meat products at a certain pH and temperature. To investigate the effects of various pH values and temperatures on the structural characteristics of Lactobacillus fermentum R6 protease, which was isolated from Harbin dry sausages, spectroscopy techniques and molecular dynamics were utilized to evaluate structural changes. RESULTS The protease exhibited a stable spatial structure at pH 7 and 40 °C, and the extension of the protease structure was also promoted. Although the structure of the protease could be changed or destroyed by pH 8 and 70 °C, it was mainly determined by the changes of secondary and tertiary structures such as α‐helix, β‐sheet, β‐turn and random coil. In addition, carbonyl vibration, ‐NH vibration, C–H stretching vibration and disulphide bonds were present in L. fermentum R6 protease under various pH and temperature conditions. Molecular docking showed that the protease can interact with myosin light chain, myosin heavy chain, actin and myoglobin. CONCLUSION The protease can maintain stable structure and interact with meat protein, which reflected certain application prospects in the fermentation of Harbin dry sausages. © 2021 Society of Chemical Industry

show abstract

Theoretical and experimental studies on the conformational changes of organic solvent-stable protease from Bacillus sphaericus DS11 in methanol/water mixtures

Cited by 31 publications

References 21 publications

Enhancement of laccase activity by pre-incubation with organic solvents

Enhancement of laccase activity by pre-incubation with organic solvents

Biochemical and Structural Characterization of Cross-Linked Enzyme Aggregates (CLEAs) of Organic Solvent Tolerant Protease

Effects of temperature and pH on the structure of a metalloprotease from Lactobacillus fermentumR6 isolated from Harbin dry sausages and molecular docking between protease and meat protein

Contact Info

Product

Resources

About