alexa Cloning and biochemical properties of a highly thermostable and enantioselective nitrilase from Alcaligenes sp. ECU0401 and its potential for (R)-(-)-mandelic acid production.
Microbiology

Microbiology

Fermentation Technology

Author(s): Zhang ZJ, Xu JH, He YC, Ouyang LM, Liu YY

Abstract Share this page

Abstract A nitrilase gene from Alcaligenes sp. ECU0401 was cloned and overexpressed in Escherichia coli BL21 (DE3) in a soluble form. The encoded protein with a His₆-tag was purified to nearly homogeneity as revealed by SDS-PAGE with a molecular weight of approximately 38.5 kDa, and the holoenzyme was estimated to be composed of 10 subunits of identical size by size exclusion chromatography. The V(max) and K(m) parameters were determined to be 27.9 μmol min⁻¹ mg⁻¹ protein and 21.8 mM, respectively, with mandelonitrile as the substrate. The purified enzyme was highly thermostable with a half life of 155 h at 30 °C and 94 h at 40 °C. Racemic mandelonitrile (50 mM) could be enantioselectively hydrolyzed to (R)-(-)-mandelic acid by the purified nitrilase with an enantiomeric excess of 97\%. The extreme stability, high activity and enantioselectivity of this nitrilase provide a solid base for its practical application in the production of (R)-(-)-mandelic acid. This article was published in Bioprocess Biosyst Eng and referenced in Fermentation Technology

Relevant Expert PPTs

Relevant Speaker PPTs

  • Ildiko Molnar
    The role of tissue-specific type 2 5’-deiodinase enzyme activities in Graves’ orbitopathy and systemic sclerosis: a new candidate in thyroid autoimmunity
    PDF Version
  • Chang-Hung Chou
    Diversified natural products in Rhododendron formosanum reveal allelochemical and pharmaceutical properties
    PPT Version | PDF Version
  • Yi-Cheng Hu
    Detection of a negative correlation between prescription of Chinese herbal products containing coumestrol, genistein or daidzein and risk of subsequent endometrial cancer among tamoxifentreated female breast cancer survivors in Taiwan between 1998 and 2008: A population-based study
    PPT Version | PDF Version
  • Carsten Worsoe
    The value of the “simulated study” as a tool to predict actual leachables in parenteral drug products
    PPT Version | PDF Version
  • Diane Paskiet
    Qualifi cation of extractables & leachables from container closure systems in drug products- Introduction to the Product Quality Institute (PQRI) recommendations
    PPT Version | PDF Version
  • Neervalur V Raghavan
    Systematic approach to development of aqueous drug formulation and drug device combination injectable products and challenges
    PPT Version | PDF Version
  • Jianfeng Hong
    Extractable and leachable studies of parenteral infusion and transfusion products
    PPT Version | PDF Version
  • Heidi Schalchli
    Natural products of Anthracophyllum discolor: Ligninolytic enzymes and antifungal volatile compounds
    PPT Version | PDF Version
  • Hazel Gorham
    Challenges in demonstrating biosimilarity and interchangeability of biosimilar products
    PPT Version | PDF Version
  • Yosef Yarden
    Classically, the 3’untranslated region (3’UTR) is that region in eukaryotic protein-coding genes from the translation termination codon to the polyA signal. It is transcribed as an integral part of the mRNA encoded by the gene. However, there exists another kind of RNA, which consists of the 3’UTR alone, without all other elements in mRNA such as 5’UTR and coding region. The importance of independent 3’UTR RNA (referred as I3’UTR) was prompted by results of artificially introducing such RNA species into malignant mammalian cells. Since 1991, we found that the middle part of the 3’UTR of the human nuclear factor for interleukin-6 (NF-IL6) or C/EBP gene exerted tumor suppression effect in vivo. Our subsequent studies showed that transfection of C/EBP 3’UTR led to down-regulation of several genes favorable for malignancy and to up-regulation of some genes favorable for phenotypic reversion. Also, it was shown that the sequences near the termini of the C/EBP 3’UTR were important for its tumor suppression activity. Then, the C/EBP 3’UTR was found to directly inhibit the phosphorylation activity of protein kinase CPKC in SMMC-7721, a hepatocarcinoma cell line. Recently, an AU-rich region in the C/EBP 3’UTR was found also to be responsible for its tumor suppression. Recently we have also found evidence that the independent C/EBP 3’UTR RNA is actually exists in human tissues, such as fetal liver and heart, pregnant uterus, senescent fibroblasts etc. Through 1990’s to 2000’s, world scientists found several 3’UTR RNAs that functioned as artificial independent RNAs in cancer cells and resulted in tumor suppression. Interestingly, majority of genes for these RNAs have promoter-like structures in their 3’UTR regions, although the existence of their transcribed products as independent 3’UTR RNAs is still to be confirmed. Our studies indicate that the independent 3’UTR RNA is a novel non-coding RNA species whose function should be the regulation not of the expression of their original mRNA, but of some essential life activities of the cell as a whole.
    PPT Version | PDF Version
  • Hyeun-Jong Bae
    Onion waste recycling to produce the value added by-products
    PPT Version | PDF Version
  • Kazuo Yano
    Regulatory approval for autologous human cells and tissue products in the United States, the European Union, and Japan
    PPT Version | PDF Version
  • Devathri Nanayakkara
    Context specific role of deubiquitylase enzyme, USP9X in oral squamous cell carcinoma
    PPT Version | PDF Version
  • Nasim Musa
    Role of angiotensin converting enzymes inhibitor and angiotensin receptor blockers in type 1 daibetic nephropathay
    PPT Version | PDF Version
  • C. Diaz
    New Antibacterial Agents against Gram-negative Pathogens from a Fundacion MEDINA’s Microbial Natural Products Collection Screening
    PPT Version | PDF Version

Recommended Conferences

Relevant Topics

Peer Reviewed Journals
 
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2017-18
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

 
© 2008-2017 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version
adwords