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Nitrilase

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https://www.readbyqxmd.com/read/28936681/production-of-r-mandelic-acid-using-nitrilase-from-recombinant-e-coli-cells-immobilized-with-tris-hydroxymethyl-phosphine
#1
Xin-Hong Zhang, Zhi-Qiang Liu, Ya-Ping Xue, Yuan-Shan Wang, Bo Yang, Yu-Guo Zheng
Recombinant Escherichia coli cells harboring nitrilase from Alcaligenes faecalis were immobilized using tris(hydroxymethyl)phosphine (THP) as the coupling agent. The optimal pH and temperature of the THP-immobilized cells were determined at pH 8.0 and 55 °C. The half-lives of THP-immobilized cells measured at 35, 40, and 50 °C were 1800, 965, and 163 h, respectively. The concentration of R-mandelic acid (R-MA) reached 358 mM after merely 1-h conversion by the immobilized cells with 500 mM R,S-mandelonitrile (R,S-MN), affording the highest productivity of 1307 g L(-1) day(-1) and the space-time productivity of 143...
September 22, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/28885614/crystal-structure-of-e-coli-apolipoprotein-n-acyl-transferase
#2
Guangyuan Lu, Yingzhi Xu, Kai Zhang, Yong Xiong, He Li, Lei Cui, Xianping Wang, Jizhong Lou, Yujia Zhai, Fei Sun, Xuejun C Zhang
In Gram-negative bacteria, lipid modification of proteins is catalysed in a three-step pathway. Apolipoprotein N-acyl transferase (Lnt) catalyses the third step in this pathway, whereby it transfers an acyl chain from a phospholipid to the amine group of the N-terminal cysteine residue of the apolipoprotein. Here, we report the 2.6-Å crystal structure of Escherichia coli Lnt. This enzyme contains an exo-membrane nitrilase domain fused to a transmembrane (TM) domain. The TM domain of Lnt contains eight TM helices which form a membrane-embedded cavity with a lateral opening and a periplasmic exit...
July 4, 2017: Nature Communications
https://www.readbyqxmd.com/read/28873512/significantly-enhanced-substrate-tolerance-of-pseudomonas-putida-nitrilase-via-atmospheric-and-room-temperature-plasma-and-cell-immobilization
#3
Ting-Ting Dong, Jin-Song Gong, Bing-Chen Gu, Qiang Zhang, Heng Li, Zhen-Ming Lu, Mao-Lin Lu, Jin-Song Shi, Zheng-Hong Xu
The objective of the study was to enhance the substrate tolerance of Pseudomonas putida nitrilase via atmospheric and room temperature plasma (ARTP) and cell immobilization. The mutant library was constructed by ARTP and rapidly screened by an OPA-TCA microscale reaction. A mutant strain of mut-D3 was obtained and its optimum substrate concentration was improved to 150mM from 100mM. It could accumulate 189g/L nicotinic acid (NA) from 3-cyanopyridine (3-CP), which was increased by 42% compared with that of wild type (WT)...
November 2017: Bioresource Technology
https://www.readbyqxmd.com/read/28858516/asymmetric-synthesis-of-akt-kinase-inhibitor-ipatasertib
#4
Chong Han, Scott Savage, Mohammad Al-Sayah, Herbert Yajima, Travis Remarchuk, Reinhard Reents, Beat Wirz, Hans Iding, Stephan Bachmann, Serena M Fantasia, Michelangelo Scalone, André Hell, Pirmin Hidber, Francis Gosselin
A highly efficient asymmetric synthesis of the Akt kinase inhibitor ipatasertib (1) is reported. The bicyclic pyrimidine 2 starting material was prepared via a nitrilase biocatalytic resolution, halogen-metal exchange/anionic cyclization, and a highly diastereoselective biocatalytic ketone reduction as key steps. The route also features a halide activated, Ru-catalyzed asymmetric hydrogenation of a vinylogous carbamic acid to produce α-aryl-β-amino acid 3 in high yield and enantioselectivity. The API was assembled in a convergent manner through a late-stage amidation/deprotection/monohydrochloride salt formation sequence...
August 31, 2017: Organic Letters
https://www.readbyqxmd.com/read/28800846/saprotrophic-proteomes-of-biotypes-of-the-witches-broom-pathogen-moniliophthora-perniciosa
#5
Sandra Pierre, Gareth W Griffith, Russell M Morphew, Luis A J Mur, Ian M Scott
Nine geographically diverse Moniliophthora perniciosa (witches' broom disease pathogen) isolates were cultured in vitro. They included six C-biotypes differing in virulence on cacao (Theobroma cacao), two S-biotypes (solanaceous hosts), and an L-biotype (liana hosts). Mycelial growth rates and morphologies differed considerably, but no characters were observed to correlate with virulence or biotype. In plant inoculations using basidiospores, one C-biotype caused symptoms on tomato (an S-biotype host), adding to evidence of limited host adaptation in these biotypes...
September 2017: Fungal Biology
https://www.readbyqxmd.com/read/28780859/photoinduced-carboborative-ring-contraction-enables-regio-and-stereoselective-synthesis-of-multiply-substituted-five-membered-carbocycles-and-heterocycles
#6
Shengfei Jin, Vu T Nguyen, Hang T Dang, Dat P Nguyen, Hadi D Arman, Oleg V Larionov
We report herein a photoinduced carboborative ring contraction of monounsaturated six-membered carbocycles and heterocycles. The reaction produces substituted five-membered ring systems stereoselectively and on preparative scales. The products feature multiple stereocenters, including contiguous quaternary carbons. We show that the reaction can serve as a synthetic platform for ring system alteration of natural products. The reaction can also be used in natural product synthesis. A concise total synthesis of artalbic acid has been enabled by a sequence of photoinduced carboborative ring contraction, Rauhut-Currier reaction, and nitrilase-catalyzed hydrolysis...
August 9, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28762035/characterization-of-a-novel-nitrilase-bgc4-from-paraburkholderia-graminis-showing-wide-spectrum-substrate-specificity-a-potential-versatile-biocatalyst-for-the-degradation-of-nitriles
#7
Haiyang Fan, Lifeng Chen, Huihui Sun, Hualei Wang, Qinghai Liu, Yuhong Ren, Dongzhi Wei
OBJECTIVE: To investigate the biodegradation of nitriles via the nitrilase-mediated pathway. RESULTS: A novel nitrilase, BGC4, was identified from proteobacteria Paraburkholderia graminis CD41M and its potential for use in biodegradation of toxic nitriles in industrial effluents was studied. BGC4 was overexpressed in Escherichia coli BL21 (DE3), the recombinant protein was purified and its enzymatic properties analysed. Maximum activity of BGC4 nitrilase was at 30 °C and pH 7...
July 31, 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/28744780/cloning-overexpression-and-characterization-of-a-thermostable-nitrilase-from-an-antarctic-pyrococcus-sp
#8
Ma Ángeles Cabrera, Jenny M Blamey
Nitriles are important chemical building blocks for the synthesis of intermediates in fine chemical and pharmaceutical industries. Here, we report a new highly thermostable nitrilase from an Antarctic Pyrococcus sp. MC-FB, a hyperthermophilic archaeon. A gene that encoded a nitrilase was identified and subsequently cloned and overexpressed in Escherichia coli. The recombinant nitrilase, named NitMC-FB, is active as a homodimer (60 kDa) with an optimal temperature and pH of 90 °C and 7.0, respectively. NitMC-FB hydrolyzes preferentially aromatic nitriles, being the first aromatic nitrilase from an archaeon described so far...
July 25, 2017: Extremophiles: Life Under Extreme Conditions
https://www.readbyqxmd.com/read/28688045/erratum-to-enhancing-the-catalytic-potential-of-nitrilase-from-pseudomonas-putida-for-stereoselective-nitrile-hydrolysis
#9
Anirban Banerjee, Praveen Kaul, U C Banerjee
No abstract text is available yet for this article.
July 8, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28623432/identification-and-expression-analysis-of-cys-a1-cys-c1-nit4-genes-in-rice-seedlings-exposed-to-cyanide
#10
Xiao-Zhang Yu, Yu-Juan Lin, Chun-Jiao Lu, Xue-Hong Zhang
Involvement of genes (CYS-A1, CYS-C1 and NIT4) encoded with cysteine synthase, β-cyanoalanine synthase, nitrilase and cyanide metabolisms are evident in Arabidopsis. In the present study, identifications of CYS-A1, CYS-C1 and NIT4, predictions of conserved motifs, and constructions of phylogenetic relationships, based on their amino acid sequences in rice, were conducted. In order to elucidate the transcriptional responses of these cyanide-degrading genes, two candidate homologues were selected for each gene to test their expression changes upon exposure to exogenous KCN in rice seedlings using RT-PCR...
June 16, 2017: Ecotoxicology
https://www.readbyqxmd.com/read/28611489/bioengineering-of-nitrilases-towards-its-use-as-green-catalyst-applications-and-perspectives
#11
REVIEW
Vinod K Nigam, Tesnim Arfi, Vishal Kumar, Pratyoosh Shukla
Nitrilases are commercial biocatalysts used for the synthesis of plastics, paints, fibers in the chemical industries, pharmaceutical drugs and herbicides for agricultural uses. Nitrilase hydrolyses the nitriles and dinitriles to their corresponding carboxylic acids and ammonia. They have a broad range of substrate specificities as well as enantio-, regio- and chemo-selective properties which make them useful for biotransformation of nitriles to important compounds because of which they are considered as 'Green Catalysts'...
June 2017: Indian Journal of Microbiology
https://www.readbyqxmd.com/read/28585068/a-novel-nitrilase-from-ralstonia-eutropha-h16-and-its-application-to-nicotinic-acid-production
#12
Haiyang Fan, Lifeng Chen, Huihui Sun, Hualei Wang, Yuhong Ren, Dongzhi Wei
A novel aliphatic nitrilase, REH16, was found in Ralstonia eutropha H16 and overexpressed in Escherichia coli BL21(DE3), and its enzymatic properties were studied. The temperature and pH optima were 37 °C and 6.6, respectively, and the best thermostability of the nitrilase was observed at 25 °C, which preserved 95% of activity after 120 h of incubation. REH16 has a broad hydrolytic activity toward aliphatic and heterocyclic nitriles and showed high tolerance of 3-cyanopyridine; this enzyme could hydrolyze as high as 100 mM 3-cyanopyridine completely...
August 2017: Bioprocess and Biosystems Engineering
https://www.readbyqxmd.com/read/28570873/biotransformation-of-nitrogen-and-sulfur-containing-pollutants-during-coking-wastewater-treatment-correspondence-of-performance-to-microbial-community-functional-structure
#13
Dev Raj Joshi, Yu Zhang, Yinxin Gao, Yuan Liu, Min Yang
Although coking wastewater is generally considered to contain high concentration of nitrogen- and sulfur-containing pollutants, the biotransformation processes of these compounds have not been well understood. Herein, a high throughput functional gene array (GeoChip 5.0) in combination with Illumina MiSeq sequencing of the 16S rRNA gene were used to identify microbial functional traits and their role in biotransformation of nitrogen- and sulfur-containing compounds in a bench-scale aerobic coking wastewater treatment system operated for 488 days...
September 15, 2017: Water Research
https://www.readbyqxmd.com/read/28498313/identifying-the-genes-regulated-by-atwrky6-using-comparative-transcript-and-proteomic-analysis-under-phosphorus-deficiency
#14
Li-Qin Li, Lu-Ping Huang, Gang Pan, Lun Liu, Xi-Yao Wang, Li-Ming Lu
Phosphorus (P) is an important mineral nutrient for plant growth and development. Overexpressing AtWRKY6 (35S:WRKY6-9) was more sensitive and wrky6 (wrky6-1) was more resistant under low Pi conditions. To better understand the function of AtWRKY6 under low phosphate stress conditions, we applied two-dimensional gel electrophoresis (2-DE) to analyse differentially expressed proteins in the shoots and roots between wild type, 35S:WRKY6-9 and wrky6-1 after phosphorus deficiency treatment for three days. The results showed 88 differentially abundant protein spots, which were identified between the shoots and roots of 35S:WRKY6-9 and wrky6-1 plants...
May 12, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28497061/mining-of-microbial-genomes-for-the-novel-sources-of-nitrilases
#15
Nikhil Sharma, Neerja Thakur, Tilak Raj, Savitri, Tek Chand Bhalla
Next-generation DNA sequencing (NGS) has made it feasible to sequence large number of microbial genomes and advancements in computational biology have opened enormous opportunities to mine genome sequence data for novel genes and enzymes or their sources. In the present communication in silico mining of microbial genomes has been carried out to find novel sources of nitrilases. The sequences selected were analyzed for homology and considered for designing motifs. The manually designed motifs based on amino acid sequences of nitrilases were used to screen 2000 microbial genomes (translated to proteomes)...
2017: BioMed Research International
https://www.readbyqxmd.com/read/28495629/improving-stability-and-reusability-of-rhodococcus-pyridinivorans-nit-36-nitrilase-by-whole-cell-immobilization-using-chitosan
#16
Jyoti, Kavita Bhatia, Kalpna Chauhan, Chandrika Attri, Amit Seth
Immobilized enzymes have great significance for industrial processes. In this study whole cell immobilization of Rhodococcus pyridinivorans NIT-36 has been undertaken using chitosan microspheres as an immobilized matrix. R. pyridinivorans NIT-36 harbors a significant intracellular enzyme nitrilase. Chitosan microspheres were generated by supplementing chitosan with glutaraldehyde and results, supported the porous microsphere structure via SEM. The resultant microspheres exhibited cell immobilization capacity of 450mg/g...
October 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28493925/a-novel-filamentous-flower-mutant-suppresses-brevipedicellus-developmental-defects-and-modulates-glucosinolate-and-auxin-levels
#17
Scott J Douglas, Baohua Li, Daniel J Kliebenstein, Eiji Nambara, C Daniel Riggs
BREVIPEDICELLUS (BP) encodes a class-I KNOTTED1-like homeobox (KNOX) transcription factor that plays a critical role in conditioning a replication competent state in the apical meristem, and it also governs growth and cellular differentiation in internodes and pedicels. To search for factors that modify BP signaling, we conducted a suppressor screen on bp er (erecta) plants and identified a mutant that ameliorates many of the pleiotropic defects of the parent line. Map based cloning and complementation studies revealed that the defect lies in the FILAMENTOUS FLOWER (FIL) gene, a member of the YABBY family of transcriptional regulators that contribute to meristem organization and function, phyllotaxy, leaf and floral organ growth and polarity, and are also known to repress KNOX gene expression...
2017: PloS One
https://www.readbyqxmd.com/read/28373563/nit1-is-a-metabolite-repair-enzyme-that-hydrolyzes-deaminated-glutathione
#18
Alessio Peracchi, Maria Veiga-da-Cunha, Tomiko Kuhara, Kenneth W Ellens, Nicole Paczia, Vincent Stroobant, Agnieszka K Seliga, Simon Marlaire, Stephane Jaisson, Guido T Bommer, Jin Sun, Kay Huebner, Carole L Linster, Arthur J L Cooper, Emile Van Schaftingen
The mammalian gene Nit1 (nitrilase-like protein 1) encodes a protein that is highly conserved in eukaryotes and is thought to act as a tumor suppressor. Despite being ∼35% sequence identical to ω-amidase (Nit2), the Nit1 protein does not hydrolyze efficiently α-ketoglutaramate (a known physiological substrate of Nit2), and its actual enzymatic function has so far remained a puzzle. In the present study, we demonstrate that both the mammalian Nit1 and its yeast ortholog are amidases highly active toward deaminated glutathione (dGSH; i...
April 18, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28286506/cyanogen-metabolism-in-cassava-roots-impact-on-protein-synthesis-and-root-development
#19
Tawanda Zidenga, Dimuth Siritunga, Richard T Sayre
Cassava (Manihot esculenta Crantz), a staple crop for millions of sub-Saharan Africans, contains high levels of cyanogenic glycosides which protect it against herbivory. However, cyanogens have also been proposed to play a role in nitrogen transport from leaves to roots. Consistent with this hypothesis, analyses of the distribution and activities of enzymes involved in cyanide metabolism provides evidence for cyanide assimilation, derived from linamarin, into amino acids in cassava roots. Both β-cyanoalanine synthase (CAS) and nitrilase (NIT), two enzymes involved in cyanide assimilation to produce asparagine, were observed to have higher activities in roots compared to leaves, consistent with their proposed role in reduced nitrogen assimilation...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28265723/cyanide-bioremediation-the-potential-of-engineered-nitrilases
#20
REVIEW
Jason M Park, B Trevor Sewell, Michael J Benedik
The cyanide-degrading nitrilases are of notable interest for their potential to remediate cyanide contaminated waste streams, especially as generated in the gold mining, pharmaceutical, and electroplating industries. This review provides a brief overview of cyanide remediation in general but with a particular focus on the cyanide-degrading nitrilases. These are of special interest as the hydrolysis reaction does not require secondary substrates or cofactors, making these enzymes particularly good candidates for industrial remediation processes...
April 2017: Applied Microbiology and Biotechnology
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