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https://www.readbyqxmd.com/read/28097501/efficient-production-of-free-fatty-acids-from-ionic-liquid-based-acid-or-enzyme-catalyzed-bamboo-hydrolysate
#1
Le Mi, Dandan Qin, Jie Cheng, Dan Wang, Sha Li, Xuetuan Wei
Two engineered Escherichia coli strains, DQ101 (MG1655 fadD (-))/pDQTES and DQ101 (MG1655 fadD (-))/pDQTESZ were constructed to investigate the free fatty acid production using ionic liquid-based acid- or enzyme-catalyzed bamboo hydrolysate as carbon source in this study. The plasmid, pDQTES, carrying an acyl-ACP thioesterase 'TesA of E. coli in pTrc99A was constructed firstly, and then (3R)-hydroxyacyl-ACP dehydratase was ligated after the TesA to give the plasmid pDQTESZ. These two strains exhibited efficient fatty acid production when glucose was used as the sole carbon source, with a final concentration of 2...
January 17, 2017: Journal of Industrial Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28089378/butyrate-production-under-aerobic-growth-conditions-by-engineered-escherichia-coli
#2
Naoya Kataoka, Alisa S Vangnai, Thunyarat Pongtharangkul, Toshiharu Yakushi, Kazunobu Matsushita
Butyrate is an important industrial platform chemical. Although several groups have reported butyrate production under oxygen-limited conditions by a native producer, Clostridium tyrobutylicum, and by a metabolically engineered Escherichia coli, efforts to produce butyrate under aerobic growth conditions have met limited success. Here, we constructed a novel butyrate synthetic pathway that functions under aerobic growth conditions in E. coli, by modifying the 1-butanol synthetic pathway reported previously...
January 11, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28071053/functional-characterization-of-the-unique-terminal-thioesterase-domain-from-polymyxin-synthetase
#3
Charles A Galea, Kade D Roberts, Yan Zhu, Philip E Thompson, Jian Li, Tony Velkov
Polymyxins remain one of the few antibiotics available for treating antibiotic resistant bacteria. Here we describe polymyxin B thioesterase which performs the final step in polymyxin B biosynthesis. Isolated thioesterase catalyzed cyclization of an N-acetylcystamine polymyxin B analogue to form polymyxin B. The thioesterase contained a catalytic cysteine unlike most thioesterases which possess a serine. Supporting this, incubation of polymyxin B thioesterase with reducing agents abolished enzymatic activity while mutation of the catalytic cysteine to serine significantly decreased activity...
January 10, 2017: Biochemistry
https://www.readbyqxmd.com/read/28065656/apt2-inhibition-restores-scribble-localization-and-s-palmitoylation-in-snail-transformed-cells
#4
Jeannie L Hernandez, Dahvid Davda, Melanie Cheung See Kit, Jaimeen D Majmudar, Sang Joon Won, Margery Gang, Sirisha C Pasupuleti, Alexandria I Choi, Callie M Bartkowiak, Brent R Martin
The multidomain scaffolding protein Scribble (Scrib) organizes key signaling complexes to specify basolateral cell polarity and suppress aberrant growth. In many human cancers, genetically normal Scrib mislocalizes from cell-cell junctions to the cytosol, correlating with enhanced growth signaling and malignancy. Here we confirm that expression of the epithelial-to-mesenchymal transcription factor (EMT-TF) Snail in benign epithelial cells leads to Scrib displacement from the plasma membrane, mimicking the mislocalization observed in aggressive cancers...
January 19, 2017: Cell Chemical Biology
https://www.readbyqxmd.com/read/28040580/physiological-effects-of-%C3%AE-linolenic-acid-and-sesamin-on-hepatic-fatty-acid-synthesis-and-oxidation
#5
Takashi Ide, Haruka Iwase, Saaya Amano, Saki Sunahara, Ayuka Tachihara, Minako Yagi, Tsuyoshi Watanabe
Interrelated effects of γ-linolenic acid (GLA) and sesamin, a sesame lignan, on hepatic fatty acid synthesis and oxidation were examined. Rats were fed experimental diets supplemented with 0 or 2 g/kg sesamin (1:1 mixture of sesamin and episesamin) and containing 100 g/kg of palm oil (saturated fat), safflower oil rich in linoleic acid, or oil of evening primrose origin containing 43% GLA (GLA oil) for 18 days. In rats fed sesamin-free diets, GLA oil, compared with other oils, increased the activity and mRNA levels of various enzymes involved in fatty acid oxidation, except for some instances...
December 14, 2016: Journal of Nutritional Biochemistry
https://www.readbyqxmd.com/read/28017601/identification-and-characterization-of-the-sulfazecin-monobactam-biosynthetic-gene-cluster
#6
Rongfeng Li, Ryan A Oliver, Craig A Townsend
The monobactams, exemplified by the natural product sulfazecin, are the only class of β-lactam antibiotics not inactivated by metallo-β-lactamases, which confer bacteria with extended-spectrum β-lactam resistance. We screened a transposon mutagenesis library from Pseudomonas acidophila ATCC 31363 and isolated a sulfazecin-deficient mutant that revealed a gene cluster encoding two non-ribosomal peptide synthetases (NRPSs), a methyltransferase, a sulfotransferase, and a dioxygenase. Three modules and an aberrant C-terminal thioesterase (TE) domain are distributed across the two NRPSs...
January 19, 2017: Cell Chemical Biology
https://www.readbyqxmd.com/read/28009004/translocation-of-the-thioesterase-domain-for-the-redesign-of-plipastatin-synthetase
#7
Ling Gao, Hongxia Liu, Zhi Ma, Jinzhi Han, Zhaoxin Lu, Chen Dai, Fengxia Lv, Xiaomei Bie
Non-ribosomal peptide synthetases (NRPSs) are large enzymatic complexes that catalyse the synthesis of biologically active peptides in microorganisms. Genetic engineering has recently been applied to reprogram NRPSs to produce lipopeptides with a new sequence. The carboxyl-terminal thioesterase (TE) domains from NRPSs catalyse cleavage products by hydrolysis or complex macrocyclization. In this study, we modified plipastatin synthetase by moving the intrinsic TE region to the end of the internal thiolation (T) domains, thus generating Bacillus subtilis strains that could produce new truncated cyclic or linear peptides of the predicted sequence, which further provided an important insight into the regioselectivity of plipastatin TE...
December 23, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27992880/a-fluorescent-probe-for-cysteine-depalmitoylation-reveals-dynamic-apt-signaling
#8
Rahul S Kathayat, Pablo D Elvira, Bryan C Dickinson
Hundreds of human proteins are modified by reversible palmitoylation of cysteine residues (S-palmitoylation), but the regulation of depalmitoylation is poorly understood. Here, we develop 'depalmitoylation probes' (DPPs), small-molecule fluorophores, to monitor the endogenous activity levels of 'erasers' of S-palmitoylation, acylprotein thioesterases (APTs). Live-cell analysis with DPPs reveals rapid growth-factor-mediated inhibition of the depalmitoylation activity of APTs, exposing a novel regulatory mechanism of dynamic lipid signaling...
February 2017: Nature Chemical Biology
https://www.readbyqxmd.com/read/27990737/tailoring-the-composition-of-novel-wax-esters-in-the-seeds-of-transgenic-camelina-sativa-through-systematic-metabolic-engineering
#9
Noemi Ruiz-Lopez, Richard Broughton, Sarah Usher, Joaquin J Salas, Richard P Haslam, Johnathan A Napier, Frédéric Beaudoin
The functional characterisation of wax biosynthetic enzymes in transgenic plants has opened the possibility of producing tailored wax esters (WEs) in the seeds of a suitable host crop. In this study, in addition to systematically evaluating a panel of WE biosynthetic activities, we have also modulated the acyl-CoA substrate pool, through the co-expression of acyl-ACP thioesterases, to direct the accumulation of medium chain fatty acids. Using this combinatorial approach, we determined the additive contribution of both the varied acyl-CoA pool and also biosynthetic enzyme substrate-specificity to the accumulation of non-native WEs in the seeds of transgenic Camelina plants...
December 19, 2016: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/27984027/generating-functional-recombinant-nrps-enzymes-in-the-laboratory-setting-via-peptidyl-carrier-protein-engineering
#10
Jeremy G Owen, Mark J Calcott, Katherine J Robins, David F Ackerley
Non-ribosomal peptide synthetases (NRPSs) are modular enzymatic assembly lines where substrates and intermediates undergo rounds of transformation catalyzed by adenylation (A), condensation (C), and thioesterase (TE) domains. Central to the NRPS biosynthesis are peptidyl carrier protein (PCP) domains, small, catalytically inactive domains that shuttle substrates and intermediates between the catalytic modules and govern product release from TE domains. There is strong interest in recombination of NRPS systems to generate new chemical entities...
November 17, 2016: Cell Chemical Biology
https://www.readbyqxmd.com/read/27983606/functional-analysis-of-the-fusarielin-biosynthetic-gene-cluster
#11
Aida Droce, Wagma Saei, Simon Hartung Jørgensen, Reinhard Wimmer, Henriette Giese, Rasmus Dam Wollenberg, Teis Esben Sondergaard, Jens Laurids Sørensen
Fusarielins are polyketides with a decalin core produced by various species of Aspergillus and Fusarium. Although the responsible gene cluster has been identified, the biosynthetic pathway remains to be elucidated. In the present study, members of the gene cluster were deleted individually in a Fusarium graminearum strain overexpressing the local transcription factor. The results suggest that a trans-acting enoyl reductase (FSL5) assists the polyketide synthase FSL1 in biosynthesis of a polyketide product, which is released by hydrolysis by a trans-acting thioesterase (FSL2)...
December 13, 2016: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/27920298/malate-synthase-and-%C3%AE-methylmalyl-coa-lyase-reactions-in-the-methylaspartate-cycle-in-haloarcula-hispanica
#12
Farshad Borjian, Jing Han, Jing Hou, Hua Xiang, Jan Zarzycki, Ivan A Berg
: Haloarchaea are extremely halophilic heterotrophic microorganisms belonging to the class Halobacteria (Euryarchaeota). Almost half of the haloarchaea possesses the genes coding for enzymes of the methylaspartate cycle, a recently discovered anaplerotic acetate assimilation pathway. In this cycle, the enzymes of the tricarboxylic acid cycle together with the dedicated enzymes of the methylaspartate cycle convert two acetyl-CoA molecules to malate. The methylaspartate cycle involves two reactions catalyzed by homologous enzymes belonging to the CitE-like enzyme superfamily, the malyl-CoA lyase/thioesterase (haloarchaeal malate synthase, hMS, Hah_2476 in Haloarcula hispanica) and the β-methylmalyl-CoA lyase (hMCL, Hah_1341)...
December 5, 2016: Journal of Bacteriology
https://www.readbyqxmd.com/read/27897367/bifunctionality-of-activ-as-a-cyclase-thioesterase-revealed-by-in-vitro-reconstitution-of-actinorhodin-biosynthesis-in-streptomyces-coelicolor-a3-2
#13
Takaaki Taguchi, Takayoshi Awakawa, Yukitaka Nishihara, Michiho Kawamura, Yasuo Ohnishi, Koji Ichinose
Type II polyketide synthase (PKS) iteratively generates a nascent polyketide thioester of acyl carrier protein (ACP), which is structurally modified to produce an ACP-free intermediate towards its final metabolite. However, the timing of ACP off-loading is not well defined due to the lack of an apparent thioesterase (TE) among relevant biosynthetic enzymes. Here, ActIV, which had been assigned as a second ring cyclase (CYC) in actinorhodin (ACT) biosynthesis, was shown to possess a TE activity in vitro with a model substrate, anthraquinone-2-carboxylic acid-N-acetylcysteamine...
November 29, 2016: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/27845206/both-a-pks-and-a-pptase-are-involved-in-melanin-biosynthesis-and-regulation-of-aureobasidium-melanogenum-xj5-1-isolated-from-the-taklimakan-desert
#14
Hong Jiang, Guang-Lei Liu, Zhe Chi, Jian-Ming Wang, Ly-Ly Zhang, Zhen-Ming Chi
A PKS1 gene responsible for the melanin biosynthesis and a NPG1 gene in Aureobasidium melanogenum XJ5-1 were cloned and characterized. An ORF of the PKS1 gene encoding a protein with 2165 amino acids contained 6495bp while an ORF of the NPG1 gene encoding a protein with 340 amino acids had 1076bp. After analysis of their promoters, it was found that expression of both the PKS1 gene and the NPG1 gene was repressed by nitrogen sources and glucose, respectively. The PKS deduced from the cloned gene consisted of one ketosynthase, one acyl transferase, two acyl carrier proteins, one thioesterase and one cyclase while the PPTase belonged to the family Sfp-type...
February 20, 2017: Gene
https://www.readbyqxmd.com/read/27765741/mri-brain-volume-measurements-in-infantile-neuronal-ceroid-lipofuscinosis
#15
E H Baker, S W Levin, Z Zhang, A B Mukherjee
BACKGROUND AND PURPOSE: Infantile neuronal ceroid lipofuscinosis is a devastating neurodegenerative storage disease caused by palmitoyl-protein thioesterase 1 deficiency, which impairs degradation of palmitoylated proteins (constituents of ceroid) by lysosomal hydrolases. Consequent lysosomal ceroid accumulation leads to neuronal injury, resulting in rapid neurodegeneration and childhood death. As part of a project studying the treatment benefits of a combination of cysteamine bitartrate and N-acetyl cysteine, we made serial measurements of patients' brain volumes with MR imaging...
October 20, 2016: AJNR. American Journal of Neuroradiology
https://www.readbyqxmd.com/read/27761152/improvement-of-free-fatty-acid-production-using-a-mutant-acyl-coa-thioesterase-i-with-high-specific-activity-in-escherichia-coli
#16
Kwang Soo Shin, Sangwoo Kim, Sung Kuk Lee
BACKGROUND: Microbial production of oleochemicals has been actively studied in the last decade. Free fatty acids (FFAs) could be converted into a variety of molecules such as industrial products, consumer products, and fuels. FFAs have been produced in metabolically engineered Escherichia coli cells expressing a signal sequence-deficient acyl-CoA thioesterase I ('TesA). Nonetheless, increasing the expression level of 'TesA seems not to be an appropriate approach to scale up FFA production because a certain ratio of each component including fatty acid synthase and 'TesA is required for optimal production of FFAs...
2016: Biotechnology for Biofuels
https://www.readbyqxmd.com/read/27748579/molecular-mechanism-for-isoform-selective-inhibition-of-acyl-protein-thioesterases-1-and-2-apt1-and-apt2
#17
Sang Joon Won, Dahvid Davda, Kristin J Labby, Sin Ye Hwang, Rachel Pricer, Jaimeen D Majmudar, Kira A Armacost, Laura A Rodriguez, Christina L Rodriguez, Fei San Chong, Kristopher A Torossian, Jasmine Palakurthi, Edward S Hur, Jennifer L Meagher, Charles L Brooks, Jeanne A Stuckey, Brent R Martin
Post-translational S-palmitoylation directs the trafficking and membrane localization of hundreds of cellular proteins, often involving a coordinated palmitoylation cycle that requires both protein acyl transferases (PATs) and acyl protein thioesterases (APTs) to actively redistribute S-palmitoylated proteins toward different cellular membrane compartments. This process is necessary for the trafficking and oncogenic signaling of S-palmitoylated Ras isoforms, and potentially many peripheral membrane proteins...
December 16, 2016: ACS Chemical Biology
https://www.readbyqxmd.com/read/27738839/functional-genomics-analysis-of-free-fatty-acid-production-under-continuous-phosphate-limiting-conditions
#18
J Tyler Youngquist, Travis C Korosh, Brian F Pfleger
Free fatty acids (FFA) are an attractive platform chemical that serves as a functional intermediate in metabolic pathways for producing oleochemicals. Many groups have established strains of Escherichia coli capable of producing various chain-length mixtures of FFA by heterologous expression of acyl-ACP thioesterases. For example, high levels of dodecanoic acid are produced by an E. coli strain expressing the Umbellularia californica FatB2 thioesterase, BTE. Prior studies achieved high dodecanoic acid yields and productivities under phosphate-limiting media conditions...
October 13, 2016: Journal of Industrial Microbiology & Biotechnology
https://www.readbyqxmd.com/read/27734184/biosynthesis-regulation-and-engineering-of-a-linear-polyketide-tautomycetin-a-novel-immunosuppressant-in-streptomyces-sp-ck4412
#19
Si-Sun Choi, Hee-Ju Nah, Hye-Rim Pyeon, Eung-Soo Kim
Tautomycetin (TMC) is a natural product with a linear structure that includes an ester bond connecting a dialkylmaleic moiety to a type I polyketide chain. Although TMC was originally identified as an antifungal antibiotic in the late 1980s, follow-up studies revealed its novel immunosuppressant activity. Specifically, TMC exhibited a mechanistically unique immunosuppressant activity about 100 times higher than that of cyclosporine A, a widely used immunosuppressant drug. Interestingly, a structurally close relative, tautomycin (TTM), was reported to not possess TMC-like immunosuppressant activity, suggesting that a distinctive polyketide moiety of TMC plays a critical role in immunosuppressant activity...
October 12, 2016: Journal of Industrial Microbiology & Biotechnology
https://www.readbyqxmd.com/read/27722792/decreased-sensitivity-of-palmitoyl-protein-thioesterase-1-deficient-neurons-to-chemical-anoxia
#20
Meredith Meyer, Attila D Kovács, David A Pearce
Infantile CLN1 disease, also known as infantile neuronal ceroid lipofuscinosis, is a fatal childhood neurodegenerative disorder caused by mutations in the CLN1 gene. CLN1 encodes a soluble lysosomal enzyme, palmitoyl protein thioesterase 1 (PPT1), and it is still unclear why neurons are selectively vulnerable to the loss of PPT1 enzyme activity in infantile CLN1 disease. To examine the effects of PPT1 deficiency on several well-defined neuronal signaling and cell death pathways, different toxic insults were applied in cerebellar granule neuron cultures prepared from wild type (WT) and palmitoyl protein thioesterase 1-deficient (Ppt1 (-/-) ) mice, a model of infantile CLN1 disease...
October 8, 2016: Metabolic Brain Disease
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