Identification and regulation of the catalytic promiscuity of (-)-γ-lactamase from Microbacterium hydrocarbonoxydans

Applied Microbiology and Biotechnology
Yu SunSheng Wu

Abstract

Mhg, a previously reported (-)-γ-lactamase from Microbacterium hydrocarbonoxydans, was identified to have perhydrolase activity by combining structure similarity search with activity assays. Kinetic studies illustrated that perhydrolysis was the native activity owing to lower K m and higher k cat/K m values. Experimental evidence showed that both hydrolysis and perhydrolysis reactions took place at the same active center. Engineering of the putative substrate-binding pocket revealed that Leu233 site played a vital role in the aspects of selective catalysis, soluble protein expression level and optimum temperature shift, etc. The mutants L233A, L233P, and L233T retained (-)-γ-lactamase activity but lost perhydrolase activity, while L233M only kept perhydrolase activity. Substitutions of Leu233 could dramatically influence the state of expressed protein. Computational analysis explicitly explained the relationships between mutations and γ-lactamase activity changes. Our investigations demonstrated that it was an efficient method to identify the enzyme catalytic promiscuity by combining 3D structure alignment with activity validations, and engineering of substrate-binding pocket could serve as a promising way to regulate activitie...Continue Reading

References

Dec 1, 1999·Bioorganic & Medicinal Chemistry·S J TaylorI N Taylor
Feb 15, 2001·Acta Crystallographica. Section D, Biological Crystallography·I S GonsalvezJ A Littlechild
Jun 22, 2001·Acta Crystallographica. Section D, Biological Crystallography·V NastopoulosD Tsernoglou
Jul 30, 2002·Journal of Molecular Biology·Beth M Beadle, Brian K Shoichet
Apr 26, 2003·Current Opinion in Chemical Biology·Shelley D Copley
Nov 6, 2003·Protein Engineering·Leila K Mosavi, Zheng-Yu Peng
Jun 24, 2004·Acta Crystallographica. Section D, Biological Crystallography·Jeremy D CheesemanRomas J Kazlauskas
Apr 2, 2005·Angewandte Chemie·Peter BernhardtRomas J Kazlauskas
Jul 20, 2005·Biochimica Et Biophysica Acta·Sharon M KellyNicholas C Price
Feb 24, 2006·Nature·Yasuo YoshikuniJay D Keasling
Oct 18, 2006·Biochemistry·Atsushi MukaiyamaShigenori Kanaya
Mar 24, 2007·Trends in Biotechnology·Karl Hult, Per Berglund
Apr 7, 2007·Journal of the American Chemical Society·Irina CatrinaDaniel Herschlag
Dec 21, 2007·Protein Engineering, Design & Selection : PEDS·Rafael CouñagoYousif Shamoo
Jan 15, 2008·Chemistry, an Asian Journal·Daniel Kahakeaw, Manfred T Reetz
Sep 27, 2008·Bioinformatics·L HolmA Schenkel
Feb 11, 2009·Nature Biotechnology·Irene NobeliJanet M Thornton
Mar 20, 2010·Annual Review of Biochemistry·Olga Khersonsky, Dan S Tawfik
May 12, 2010·Nucleic Acids Research·Liisa Holm, Päivi Rosenström
Jun 17, 2010·Bioinformatics·Pablo Carbonell, Jean-Loup Faulon
Oct 30, 2012·Journal of the American Chemical Society·Dan Thomas Major, Michal Weitman
Feb 27, 2014·Bioorganic & Medicinal Chemistry Letters·Hongtao Zhao, Amedeo Caflisch
May 23, 2014·Chembiochem : a European Journal of Chemical Biology·Catherine B PoorJared C Lewis
Dec 17, 2014·Applied Microbiology and Biotechnology·Jianjun WangSheng Wu

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.