Site-directed mutation to improve the enzymatic activity of 5-carboxy-2-pentenoyl-CoA reductase for enhancing adipic acid biosynthesis

2019 ◽  
Vol 125 ◽  
pp. 6-12 ◽  
Author(s):  
Ju Yang ◽  
Yiting Lu ◽  
Yunying Zhao ◽  
Zhonghu Bai ◽  
Zheng Ma ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Adipic Acid ◽  
Acid Biosynthesis ◽  
Directed Mutation ◽  
Coa Reductase

scholarly journals Biosynthesis ofcis,cis-Muconic Acid and Its Aromatic Precursors, Catechol and Protocatechuic Acid, from Renewable Feedstocks by Saccharomyces cerevisiae

2012 ◽  
Vol 78 (23) ◽  
pp. 8421-8430 ◽  
Author(s):  
Christian Weber ◽  
Christine Brückner ◽  
Sheila Weinreb ◽  
Claudia Lehr ◽  
Christine Essl ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Amino Acid ◽  
Adipic Acid ◽  
Aromatic Amino Acid ◽  
Protocatechuic Acid ◽  
Amino Acid Biosynthesis ◽  
Biosynthesis Pathway ◽  
Acid Biosynthesis ◽  
Muconic Acid ◽  
Production Processes

ABSTRACTAdipic acid is a high-value compound used primarily as a precursor for the synthesis of nylon, coatings, and plastics. Today it is produced mainly in chemical processes from petrochemicals like benzene. Because of the strong environmental impact of the production processes and the dependence on fossil resources, biotechnological production processes would provide an interesting alternative. Here we describe the first engineeredSaccharomyces cerevisiaestrain expressing a heterologous biosynthetic pathway converting the intermediate 3-dehydroshikimate of the aromatic amino acid biosynthesis pathway via protocatechuic acid and catechol intocis,cis-muconic acid, which can be chemically dehydrogenated to adipic acid. The pathway consists of three heterologous microbial enzymes, 3-dehydroshikimate dehydratase, protocatechuic acid decarboxylase composed of three different subunits, and catechol 1,2-dioxygenase. For each heterologous reaction step, we analyzed several potential candidates for their expression and activity in yeast to compose a functionalcis,cis-muconic acid synthesis pathway. Carbon flow into the heterologous pathway was optimized by increasing the flux through selected steps of the common aromatic amino acid biosynthesis pathway and by blocking the conversion of 3-dehydroshikimate into shikimate. The recombinant yeast cells finally produced about 1.56 mg/litercis,cis-muconic acid.

The role of symmetry in the regulation of bacterial carboxyltransferase

2011 ◽  
Vol 2 (1-2) ◽  
pp. 47-52 ◽  
Author(s):  
Grover L. Waldrop
Keyword(s):  
Gene Expression ◽  
Enzymatic Activity ◽  
Chemical Reaction ◽  
Binding Sites ◽  
Fatty Acid Biosynthesis ◽  
Acid Biosynthesis ◽  
Effective Mechanism ◽  
Acetyl Coa ◽  
Mrna Binding

AbstractCarboxyltransferase is one component of the multifunctional enzyme acetyl-CoA carboxylase which catalyzes the first committed step in fatty acid biosynthesis. Carboxyltransferase is an α2β2heterotetramer and possesses two distinct but integrated functions. One function catalyzes the transfer of carbon dioxide from biotin to acetyl-CoA, whereas the other involves binding to the mRNA encoding both subunits. When carboxyltransferase binds to the mRNA both enzymatic activity and translation of the mRNA are inhibited. However, the substrate acetyl-CoA competes with mRNA for binding. Thus, mRNA binding by carboxyltransferase provides an effective mechanism for regulating enzymatic activity and gene expression. This conceptual review takes the position that regulation of enzymatic activity and gene expression of carboxyltransferase by binding to its own mRNA is at its most fundamental level the result of the symmetry in the chemical reaction catalyzed by the enzyme. The chemical reaction is symmetrical in that both substrates generate enolate anions during the course of catalysis. The chemical symmetry led to a structural symmetry in the enzyme where both the α and β subunits contain oxyanion holes that stabilize the enolate anions. Then the region of the mRNA that codes for the oxyanion holes provided the binding sites for carboxyltransferase. Thus, the symmetry of the chemical reaction formed the foundation for the evolution of the mechanism for regulation of carboxyltransferase.

scholarly journals Exploring functionality of the reverse β-oxidation pathway in Corynebacterium glutamicum for production of adipic acid

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Jae Ho Shin ◽  
Aaron John Christian Andersen ◽  
Puck Achterberg ◽  
Lisbeth Olsson
Keyword(s):  
Corynebacterium Glutamicum ◽  
Adipic Acid ◽  
Condensation Product ◽  
Tca Cycle ◽  
Cell Factory ◽  
Acetyl Coa ◽  
Oxidation Pathway ◽  
Model Cell ◽  
Coa Reductase ◽  
Cultivation Broth

Abstract Background Adipic acid, a six-carbon platform chemical mainly used in nylon production, can be produced via reverse β-oxidation in microbial systems. The advantages posed by Corynebacterium glutamicum as a model cell factory for implementing the pathway include: (1) availability of genetic tools, (2) excretion of succinate and acetate when the TCA cycle becomes overflown, (3) initiation of biosynthesis with succinyl-CoA and acetyl-CoA, and (4) established succinic acid production. Here, we implemented the reverse β-oxidation pathway in C. glutamicum and assessed its functionality for adipic acid biosynthesis. Results To obtain a non-decarboxylative condensation product of acetyl-CoA and succinyl-CoA, and to subsequently remove CoA from the condensation product, we introduced heterologous 3-oxoadipyl-CoA thiolase and acyl-CoA thioesterase into C. glutamicum. No 3-oxoadipic acid could be detected in the cultivation broth, possibly due to its endogenous catabolism. To successfully biosynthesize and secrete 3-hydroxyadipic acid, 3-hydroxyadipyl-CoA dehydrogenase was introduced. Addition of 2,3-dehydroadipyl-CoA hydratase led to biosynthesis and excretion of trans-2-hexenedioic acid. Finally, trans-2-enoyl-CoA reductase was inserted to yield 37 µg/L of adipic acid. Conclusions In the present study, we engineered the reverse β-oxidation pathway in C. glutamicum and assessed its potential for producing adipic acid from glucose as starting material. The presence of adipic acid, albeit small amount, in the cultivation broth indicated that the synthetic genes were expressed and functional. Moreover, 2,3-dehydroadipyl-CoA hydratase and β-ketoadipyl-CoA thiolase were determined as potential target for further improvement of the pathway.

scholarly journals Atorvastatin Improves Hepatic Lipid Metabolism and Protects Renal Damage in Adenine-Induced Chronic Kidney Disease in Sprague-Dawley Rats

2019 ◽  
Vol 2019 ◽  
pp. 1-10
Author(s):  
Hardik Ghelani ◽  
Valentina Razmovski-Naumovski ◽  
Vamsi Inampudi ◽  
Dennis Chang ◽  
Srinivas Nammi
Keyword(s):  
Lipid Metabolism ◽  
Kidney Disease ◽  
Enzymatic Activity ◽  
Cholesterol Acyltransferase ◽  
Ldl Receptor ◽  
Hepatic Lipid Metabolism ◽  
Hmg Coa Reductase ◽  
Hepatic Lipid ◽  
Atorvastatin Therapy ◽  
Coa Reductase

Objective. Chronic kidney disease (CKD), including nephrotic syndrome, is a major cause of cardiovascular morbidity and mortality. The literature indicates that CKD is associated with profound lipid disorders largely due to the dysregulation of lipoprotein metabolism which further aggravates the progression of kidney disease. The present study sought to determine the efficacy of atorvastatin treatment on hepatic lipid metabolism and renal tissue damage in CKD rats. Methods. Serum, hepatic and faecal lipid contents and the expression and enzyme activity of molecules involved in cholesterol and triglyceride metabolism, along with kidney function, were determined in untreated adenine-induced CKD, atorvastatin-treated CKD (10 mg/kg/day oral for 24 days) and control rats. Key Findings. CKD resulted in metabolic dyslipidaemia, renal insufficiency, hepatic lipid accumulation, upregulation of 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase, acyl-CoA cholesterol acyltransferase-2 (ACAT2) and the downregulation of LDL receptor protein, VLDL receptor, hepatic lipase, lipoprotein lipase (LPL), lecithin–cholesterol acyltransferase (LCAT) and scavenger receptor class B type 1 (SR-B1). CKD also resulted in increased enzymatic activity of HMG-CoA reductase and ACAT2 together with decreased enzyme activity of lipase and LCAT. Atorvastatin therapy attenuated dyslipidaemia, renal insufficiency, reduced hepatic lipids, HMG-CoA reductase and ACAT2 protein abundance and raised LDL receptor and lipase protein expression. Atorvastatin therapy decreased the enzymatic activity of HMG-CoA reductase and increased enzymatic activity of lipase and LCAT. Conclusions. Atorvastatin improved hepatic tissue lipid metabolism and renal function in adenine-induced CKD rats.

A Multiperspective Approach to Solvent Regulation of Enzymatic Activity: HMG-CoA Reductase

ChemBioChem ◽  
2017 ◽  
Vol 19 (2) ◽  
pp. 153-158 ◽  
Author(s):  
Michael Dirkmann ◽  
Javier Iglesias-Fernández ◽  
Victor Muñoz ◽  
Pandian Sokkar ◽  
Christoph Rumancev ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Hmg Coa Reductase ◽  
Coa Reductase

Directed mutation of β-glucanases from probiotics to enhance enzymatic activity, thermal and pH stability

2020 ◽  
Vol 202 (7) ◽  
pp. 1749-1756
Author(s):  
Zhan-Bin Sun ◽  
Jia-Liang Xu ◽  
Xin Lu ◽  
Wei Zhang ◽  
Chao Ji ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Ph Stability ◽  
Directed Mutation

scholarly journals Stimulatory Effects of Oleci Acid and Fungal Elicitor on Betulinic Acid Production by Submerged Cultivation of Medicinal Mushroom Inonotus obliquus

2021 ◽  
Vol 7 (4) ◽  
pp. 266
Author(s):  
Hanghang Lou ◽  
Hao Li ◽  
Tianyu Wei ◽  
Qihe Chen
Keyword(s):  
Oleic Acid ◽  
Betulinic Acid ◽  
Fermentation Broth ◽  
Fungal Elicitor ◽  
Submerged Cultivation ◽  
Medicinal Mushroom ◽  
Acid Biosynthesis ◽  
Inonotus Obliquus ◽  
Key Genes ◽  
Coa Reductase

To evaluate the novel strategy of oleic acid and fungal elicitor (made from Aspergillus niger) to elicit betulinic acid biosynthesis in medicinal mushroom Inonotus obliquus, we conduct the stimulatory effects investigation for synthesizing betulinic acid from betulin. HPLC results indicated oleic acid and fungal elicitor were effective stimulators. The supplementation of 1.0 g/L oleic acid led to the highest increase of betulinic acid either in dry mycelia or fermentation broth by 2-fold of the control. Fungal elicitor at 45 mg/L markedly increases mycelia growth by 146.0% and enhance intracellular betulinic acid accumulation by 429.5% as compared to the controls. Quantification of transcription levels determined that oleic acid, fungal elicitor and their combinations could induce the expressions of key genes involved in betulinic acid biosynthesis, such as HMG-CoA reductase and squalene synthase. These findings indicated that oleic acid and fungal elicitor could enhance betulinic acid metabolism by up-regulating key genes expression.

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