scholarly journals Enhancing the Methanol Tolerance of Candida antarctica Lipase B by Saturation Mutagenesis for Biodiesel Preparation

2021 ◽  
Author(s):  
Zhongbiao Tan ◽  
Xiangqian Li ◽  
Hao Shi ◽  
Xiulian Yin ◽  
Xiaoyan Zhu ◽  
...  
Keyword(s):  
Candida Antarctica ◽  
Molecular Engineering ◽  
Saturation Mutagenesis ◽  
Methanol Tolerance ◽  
Lipase B ◽  
Hydrophobic Force ◽  
Factors Affecting ◽  
Solid Plate ◽  
Methanol Tolerant ◽  
Formed Hydrogen

Abstract Methanol tolerance of a lipase is one of the important factors affecting its esterification ability in biodiesel preparation. By B factor indicated prediction of Candida antarctica lipase B (CalB) surface amino acids, 8 sites (Val139, Ala146, Leu147, Pro218, Val286, Ala287, Val306, and Gly307) with high B value indicating more flexibility were chosen to perform saturation mutagenesis. High-methanol-tolerant variants, CalB-P218W and -V306N, created larger haloes on emulsified tributyrin solid plate including 15% (v/v) methanol and showed 19% and 31% higher activity over CalB-WT (wild type), respectively. By modeling, a newly formed hydrogen bond in CalB-V306N and hydrophobic force in CalB-P218W contributing more stability in protein may have resulted in increased methanol tolerance. CalB-P218W and -V306N transesterified the soybean oil into biodiesel at 30 °C by 85% and 89% yield, respectively, over 82% by CalB-WT for 24 h reactions. These results may provide a basis for molecular engineering of CalB and expand its applications in fuel industries. The as-developed semi-rational method could be utilized to enhance the stabilities of many other industrial enzymes.

scholarly journals Influence of Glutaraldehyde Cross-Linking Modes on the Recyclability of Immobilized Lipase B from Candida antarctica for Transesterification of Soy Bean Oil

Molecules ◽  
2018 ◽  
Vol 23 (9) ◽  
pp. 2230 ◽  
Author(s):  
Iago A. Modenez ◽  
Diego Sastre ◽  
Fernando C. Moraes ◽  
Caterina Marques Netto
Keyword(s):  
Magnetic Nanoparticles ◽  
Immobilized Lipase ◽  
Carbon Chain ◽  
Candida Antarctica ◽  
Cross Linking ◽  
Carbon Chain Length ◽  
Lipase B ◽  
Factors Affecting ◽  
Michael Type Addition ◽  
Nanoparticle Agglomerate

Lipase B from Candida antarctica (CAL-B) is largely employed as a biocatalyst for hydrolysis, esterification, and transesterification reactions. CAL-B is a good model enzyme to study factors affecting the enzymatic structure, activity and/or stability after an immobilization process. In this study, we analyzed the immobilization of CAL-B enzyme on different magnetic nanoparticles, synthesized by the coprecipitation method inside inverse micelles made of zwitterionic surfactants, with distinct carbon chain length: 4 (ImS4), 10 (ImS10) and 18 (ImS18) carbons. Magnetic nanoparticles ImS4 and ImS10 were shown to cross-link to CAL-B enzyme via a Michael-type addition, whereas particles with ImS18 were bond via pyridine formation after glutaraldehyde cross-coupling. Interestingly, the Michael-type cross-linking generated less stable immobilized CAL-B, revealing the influence of a cross-linking mode on the resulting biocatalyst behavior. Curiously, a direct correlation between nanoparticle agglomerate sizes and CAL-B enzyme reuse stability was observed. Moreover, free CAL-B enzyme was not able to catalyze transesterification due to the high methanol concentration; however, the immobilized CAL-B enzyme reached yields from 79.7 to 90% at the same conditions. In addition, the transesterification of lipids isolated from oleaginous yeasts achieved 89% yield, which confirmed the potential of immobilized CAL-B enzyme in microbial production of biodiesel.

scholarly journals New Efficient Recombinant Expression System To Engineer Candida antarctica Lipase B

2010 ◽  
Vol 76 (8) ◽  
pp. 2684-2687 ◽  
Author(s):  
St�phane Emond ◽  
C�dric Montanier ◽  
Jean-Marc Nicaud ◽  
Alain Marty ◽  
Pierre Monsan ◽  
...  
Keyword(s):  
Recombinant Expression ◽  
Expression System ◽  
Candida Antarctica ◽  
Saturation Mutagenesis ◽  
Wild Type ◽  
Wild Type Enzyme ◽  
Lipase B ◽  
Expression Host ◽  
Screening Protocol ◽  
Recombinant Expression System

ABSTRACT Here, we report the use of Yarrowia lipolytica as a versatile expression host for developing protein engineering approaches to modify the properties of Candida antarctica lipase B. A reliable screening protocol was defined and validated using a saturation mutagenesis library, yielding mutants displaying higher catalytic efficiencies than the wild-type enzyme.

Factors Affecting Molecular Self-Assembly and Its Mechanism

2012 ◽  
Vol 9 (1) ◽  
pp. 43 ◽  
Author(s):  
Hueyling Tan
Keyword(s):  
Self Assembly ◽  
Building Blocks ◽  
Molecular Engineering ◽  
Molecular Structures ◽  
Polymer Science ◽  
New Approach ◽  
Factors Affecting ◽  
Dna Structures ◽  
Self Assembling ◽  
Wide Range

Molecular self-assembly is ubiquitous in nature and has emerged as a new approach to produce new materials in chemistry, engineering, nanotechnology, polymer science and materials. Molecular self-assembly has been attracting increasing interest from the scientific community in recent years due to its importance in understanding biology and a variety of diseases at the molecular level. In the last few years, considerable advances have been made in the use ofpeptides as building blocks to produce biological materials for wide range of applications, including fabricating novel supra-molecular structures and scaffolding for tissue repair. The study ofbiological self-assembly systems represents a significant advancement in molecular engineering and is a rapidly growing scientific and engineering field that crosses the boundaries ofexisting disciplines. Many self-assembling systems are rangefrom bi- andtri-block copolymers to DNA structures as well as simple and complex proteins andpeptides. The ultimate goal is to harness molecular self-assembly such that design andcontrol ofbottom-up processes is achieved thereby enabling exploitation of structures developed at the meso- and macro-scopic scale for the purposes oflife and non-life science applications. Such aspirations can be achievedthrough understanding thefundamental principles behind the selforganisation and self-synthesis processes exhibited by biological systems.

Modelling and Predicting Enzyme Enantioselectivity: the Aid of Computational Methods for the Rational use of Lipase B from Candida antarctica

2015 ◽  
Vol 4 (2) ◽  
pp. 87-99 ◽  
Author(s):  
Valerio Ferrario ◽  
Cynthia Ebert ◽  
Patrizia Nitti ◽  
Giuliana Pitacco ◽  
Lucia Gardossi
Keyword(s):  
Computational Methods ◽  
Candida Antarctica ◽  
Lipase B ◽  
Rational Use

The effect of decationation of hydroxylated HNaY zeolites on their interaction with ethylene and propylene. hydrogen complexes, oligomeration and isotope exchange

1980 ◽  
Vol 45 (11) ◽  
pp. 3101-3115 ◽  
Author(s):  
Ludmila Kubelková ◽  
Jana Nováková ◽  
Zdeněk Dolejšek ◽  
Pavel Jírů
Keyword(s):  
Isotope Exchange ◽  
Decisive Role ◽  
Single Step ◽  
Hydroxyl Groups ◽  
Isotope Exchange Reaction ◽  
Factors Affecting ◽  
Step Mechanism ◽  
Formed Hydrogen ◽  
Lower Basicity ◽  
Multiple Step

The effect of decationation on the interaction of propylene and ethylene with the hydroxylated forms of HNaY zeolites has been studied. The compounds formed in the zeolite cavities were studied using their infrared spectra, the composition of the gaseous phase was followed by mass spectrometry. The results showed that among factors affecting the interaction with propylene properties of hydroxyl groups play the decisive role. With the increasing decationation of the zeolite the strength of the OH bond in the hydrogen complex of the large-cavity hydroxyls with propylene decreased and the rate of propylene oligomeration at 310 K as well as the isotope exchange rate of propylene-d6 with the zeolite hydroxyls at 570 K increased. The propylene isotope exchange reaction proceeded by the multiple-step mechanism. In the case of ethylene only the physical sorption with a non-specific character was observed at 310 K. The adsorbed amount increased with the increasing content of Na+ ions in the zeolite (with the decreasing decationation). A part of the adsorbed molecules formed hydrogen complexes with hydroxyls of large cavities. The observed lower weakening of the OH bond was in agreement with the lower basicity of ethylene if compared with propylene. The isotope exchange of ethylene-d4 with the zeolite hydroxyls proceeded by a single-step mechanism, it had an autocatalytic character and its rate was 70 times lower than that of the isotope exchange of propylene-d6 with OH.

Pore-expanded SBA-15 for the immobilization of a recombinant Candida antarctica lipase B: Application in esterification and hydrolysis as model reactions

2018 ◽  
Vol 129 ◽  
pp. 12-24 ◽  
Author(s):  
Nathalia Saraiva Rios ◽  
Maisa Pessoa Pinheiro ◽  
Magno Luís Bezerra Lima ◽  
Denise Maria Guimarães Freire ◽  
Ivanildo José da Silva ◽  
...  
Keyword(s):  
Candida Antarctica ◽  
Candida Antarctica Lipase B ◽  
Candida Antarctica Lipase ◽  
Lipase B ◽  
Model Reactions

Structural effects on chemo- and enantioselectivity of Candida antarctica lipase B - Resolution of β-amino esters

2002 ◽  
Vol 80 (6) ◽  
pp. 565-570 ◽  
Author(s):  
Szilvia Gedey ◽  
Arto Liljeblad ◽  
László Lázár ◽  
Ferenc Fülöp ◽  
Liisa T Kanerva
Keyword(s):  
Key Words ◽  
Candida Antarctica ◽  
Diisopropyl Ether ◽  
The Other ◽  
Candida Antarctica Lipase B ◽  
Candida Antarctica Lipase ◽  
Structural Effects ◽  
Lipase B ◽  
Amino Esters ◽  
Catalyzed Reactions

The Candida antarctica lipase B-catalyzed reactions of five β-amino esters with neat butyl butanoate and with 2,2,2-trifluoroethyl butanoate in diisopropyl ether were studied, as were the reactions of the same β-amino esters and their N-butanamides with neat butanol. The possibility for sequential resolution, where the amino and ester functions of the substrate both react with an achiral butanoate, became less likely with increasing size of the substrate from ethyl 3-aminobutanoate (1a) to pentanoate (1b) or larger. On the other hand, the alcoholyses of N-acylated β-amino esters successfully proceeded in butanol with E > 100. Gram-scale resolution of the N-butanoylated 1a was performed to demonstrate the usefulness of the method. Key words: lipase, interesterification, acylation, alcoholysis, resolution, β-amino esters.

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