Lipolytic enzymes that retain high levels of catalytic activity when exposed to a variety of denaturing conditions are of high importance for a number of biotechnological applications. In this study, we aimed to identify new lipolytic enzymes, which are highly resistant to prolonged exposure to elevated temperatures. To achieve this, we searched for genes encoding for such proteins in the genomes of a microbial consortium residing in a hot spring located in China. After performing functional genomic screening on a bacterium of the genus , which was isolated from this hot spring following enrichment, we identified a new esterolytic enzyme, termed EstDZ3. Detailed biochemical characterization of the recombinant enzyme, revealed that it constitutes a slightly alkalophilic and highly active esterase against esters of fatty acids with short to medium chain lengths. Importantly, EstDZ3 exhibits remarkable thermostability, as it retains high levels of catalytic activity after exposure to temperatures as high as 95°C for several hours. Furthermore, it exhibits very good stability against exposure to high concentrations of a variety of organic solvents. Interestingly, EstDZ3 was found to have very little similarity to previously characterized esterolytic enzymes. Computational modeling of the three-dimensional structure of this new enzyme predicted that it exhibits a typical α/β hydrolase fold that seems to include a "subdomain insertion", which is similar to the one present in its closest homolog of known function and structure, the cinnamoyl esterase Lj0536 from . As it was found in the case of Lj0536, this structural feature is expected to be an important determinant of the catalytic properties of EstDZ3. The high levels of esterolytic activity of EstDZ3, combined with its remarkable thermostability and good stability against a range of organic solvents and other denaturing agents, render this new enzyme a candidate biocatalyst for high-temperature biotechnological applications.
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http://dx.doi.org/10.3389/fmicb.2016.01779 | DOI Listing |
Int J Biol Macromol
January 2025
College of Life Science, Northwest A&F University, Yangling 712100, China. Electronic address:
Amycolatopsis sp. BJA-103 was isolated for its exceptional feather-degradation capability, leading to the purification, cloning, and heterologous expression of the keratinase enzyme, KER0199. Sequence analysis places KER0199 within the S8 protease family, revealing <60 % sequence similarity to known proteases.
View Article and Find Full Text PDFInt J Biol Macromol
December 2024
College of Materials Science and Engineering, Nanjing Tech University, Nanjing 211816, Jiangsu Province, China. Electronic address:
Over the past decades, emerging bioplastics have attracted much interest from the scientific and industrial communities because of public concerns about environmental problems and sustainable development. In this study, poly(lactic acid) (PLA) was toughened by ductile biodegradable poly(butylene adipate-co-terephthalate) (PBAT) and biosourced plasticizer epoxidized linseed oil (ELO), and a chain-extending agent (CEA) was added to promote the compatibility and toughness of the bio-blends. It was shown that "in situ" grafted polymers were created in the bio-blends with the aid of CEA, greatly enhancing the compatibility and ductility of the compatibilized blends.
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December 2024
Department of Molecular Cell and Developmental Biology, University of California, Santa Cruz, California, USA; Center for Molecular Biology of RNA, University of California, Santa Cruz, California, USA. Electronic address:
The spliceosome protein, SF3B1 associates with U2 snRNP during early spliceosome assembly for pre-mRNA splicing. Frequent somatic mutations in SF3B1 observed in cancer necessitates characterization of its role in identifying the branchpoint adenosine of introns. Remarkably, SF3B1 is the target of three distinct natural product drugs, each identified by their potent anti-tumor properties.
View Article and Find Full Text PDFJ Agric Food Chem
December 2024
Key Laboratory of Industrial Biotechnology of the Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
In addressing the challenges posed by extended fermentation cycles and high-salt conditions in high-salt liquid-state fermentation soy sauce (HLFSS) production, a high-throughput screening method was devised to identify thermally stable l-glutaminase mutants. This study yielded mutants A146D and A51D, exhibiting enhanced thermal stability. Computer-aided analysis revealed that these mutations introduced additional forces, compacting the protein structure and lowering the Gibbs free energy, thereby improving thermostability.
View Article and Find Full Text PDFProc Natl Acad Sci U S A
December 2024
State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
DNA repair systems are essential to maintain genome integrity and stability. Some obligate endosymbionts that experience long-term symbiosis with the insect hosts, however, have lost their key components for DNA repair. It is largely unexplored how the bacterial endosymbionts cope with the increased demand for mismatch repairs under heat stresses.
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