Exploring the Protein Stability Landscape: <i>Bacillus subtilis</i> Lipase A as a Model for Detergent Tolerance

Fulton, Alexander; Frauenkron-Machedjou, Victorine Josiane; Skoczinski, Pia; Wilhelm, Susanne; Zhu, Leilei; Schwaneberg, Ulrich; Jaeger, Karl‐Erich

doi:10.1002/cbic.201402664

Cited by 46 publications

(55 citation statements)

References 45 publications

(41 reference statements)

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“…Microbial lipases specifically have unlimited potential in commercial applications, such as additives in fine chemicals, wastewater treatment, food processing, cosmetics, detergents, pharmaceuticals, degreasing formulations, paper manufacture, and accelerated degradation of fatty wastes and polyurethane (Rodriques et al 2016;Shu et al 2016;Speranza et al 2016;Fulton et al 2015;Adulkar and Rathod 2014;Gerits et al 2014;Li et al 2014;Saranya et al 2014;Brabcova et al 2013;Lailaja and Chandrasekaran 2013;Nerurkar et al 2013;Whangsuk et al 2013;Zhang et al 2013;Liu et al 2012;Kamini et al 2000). Even though a large number of lipases have been described in the literature, only for a limited number of enzymes belonging to a few species has it been proven that their stability and biosynthetic activity is amenable for use in organic solvents, and thus for consideration as industrially applicable enzymes (Cardenas et al 2001;Jaeger et al 1999).…”

Section: Lipasesmentioning

confidence: 99%

Classification of lipolytic enzymes and their biotechnological applications in the pulping industry

2017

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Abstract:In the pulp and paper industry, during the manufacturing process, the agglomeration of pitch particles (composed of triglycerides, fatty acids, and esters) leads to the formation of black pitch deposits in the pulp and on machinery, which impacts on the process and pulp quality. Traditional methods of pitch prevention and treatment are no longer feasible due to environmental impact and cost. Consequently, there is a need for more efficient and environmentally friendly approaches. The application of lipolytic enzymes, such as lipases and esterases, could be the sustainable solution to this problem. Therefore, an understanding of their structure, mechanism, and sources are essential. In this report, we review the microbial sources for the different groups of lipolytic enzymes, the differences between lipases and esterases, and their potential applications in the pulping industry.

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Section: Lipasesmentioning

confidence: 99%

Classification of lipolytic enzymes and their biotechnological applications in the pulping industry

2017

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“…An SSM library covering all natural diversity with a single amino acid exchange (named 'BSLA-SSM library') has been previously reported Fulton et al 2015). The BSLA-SSM library mimics natural evolution, which evolves enzymes by introducing single amino acid exchanges due to its very low mutation frequency (Lynch 2010).…”

Section: Introductionmentioning

confidence: 99%

Exploring the full natural diversity of single amino acid exchange reveals that 40–60% of BSLA positions improve organic solvents resistance

Frauenkron-Machedjou

Fulton

Zhao

et al. 2018

Bioresour. Bioprocess.

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Objectives: Protein engineering has been employed to successfully improve organic solvent resistance of enzymes. Exploration of nature's full potential (how many beneficial positions/beneficial substitutions of the target enzyme) to improve organic solvent resistance of enzymes by a systematic study was performed. Results:We report the results of screening the previously generated BSLA (Bacillus subtilis lipase A)-SSM (site saturation mutagenesis) library (covering the full natural diversity of BSLA with one amino acid exchange) in presence of three cosolvents. The potential of single amino acid substitution that nature offers to improve the cosolvent resistance of BSLA was determined by analyzing the number of beneficial positions/substitutions, accessibility and chemical compositions. Conclusion:Lessons learned from analysis of BSLA-SSM library are: (1) 41-59% of BSLA positions with in total 4-10% of all possible substitutions improve the cosolvent resistance against TFE, DOx, and DMSO; (2) charged substitutions are preferred to improve DOx and TFE resistance whereas polar ones are preferred for DMSO; (3) charged substitutions on the surface predominantly improved resistance while polar ones were preferred in buried "regions". (4) Interestingly, 58-93% of beneficial substitutions led to chemically different amino acids.

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“…An important strategy to guide enzyme‐engineering efforts is to make use of mutability landscapes . By screening a large collection of nearly all single mutants of an enzyme, important information is obtained on single mutations or residue positions that influence a desired characteristic of the enzyme.…”

Section: Discussionmentioning

confidence: 99%

Tuning Enzyme Activity for Nonaqueous Solvents: Engineering an Enantioselective “Michaelase” for Catalysis in High Concentrations of Ethanol

et al. 2020

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Enzymes have evolved to function under aqueous conditions and may not exhibit features essential for biocatalytic application, such as the ability to function in high concentrations of an organic solvent. Consequently, protein engineering is often required to tune an enzyme for catalysis in non‐aqueous solvents. In this study, we have used a collection of nearly all single mutants of 4‐oxalocrotonate tautomerase, which promiscuously catalyzes synthetically useful Michael‐type additions of acetaldehyde to various nitroolefins, to investigate the effect of each mutation on the ability of this enzyme to retain its “Michaelase” activity in elevated concentrations of ethanol. Examination of this mutability landscape allowed the identification of two hotspot positions, Ser30 and Ala33, at which mutations are beneficial for catalysis in high ethanol concentrations. The “hotspot” position Ala33 was then randomized in a highly enantioselective, but ethanol‐sensitive 4‐OT variant (L8F/M45Y/F50A) to generate an improved enzyme variant (L8F/A33I/M45Y/F50A) that showed great ethanol stability and efficiently catalyzes the enantioselective addition of acetaldehyde to nitrostyrene in 40 % ethanol (permitting high substrate loading) to give the desired γ‐nitroaldehyde product in excellent isolated yield (89 %) and enantiopurity (ee=98 %). The presented work demonstrates the power of mutability‐landscape‐guided enzyme engineering for efficient biocatalysis in non‐aqueous solvents.

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Exploring the Protein Stability Landscape: Bacillus subtilis Lipase A as a Model for Detergent Tolerance

Cited by 46 publications

References 45 publications

Classification of lipolytic enzymes and their biotechnological applications in the pulping industry

Classification of lipolytic enzymes and their biotechnological applications in the pulping industry

Exploring the full natural diversity of single amino acid exchange reveals that 40–60% of BSLA positions improve organic solvents resistance

Tuning Enzyme Activity for Nonaqueous Solvents: Engineering an Enantioselective “Michaelase” for Catalysis in High Concentrations of Ethanol

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