Asymmetric reductive amination by a wild-type amine dehydrogenase from the thermophilic bacteria Petrotoga mobilis

2016 ◽  
Vol 6 (20) ◽  
pp. 7421-7428 ◽  
Author(s):  
Ombeline Mayol ◽  
Sylvain David ◽  
Ekaterina Darii ◽  
Adrien Debard ◽  
Aline Mariage ◽  
...  
Keyword(s):  
Primary Amine ◽  
Reductive Amination ◽  
Thermophilic Bacteria ◽  
Wild Type ◽  
Enantiomerically Pure ◽  
Amine Dehydrogenase

Biocatalytic potential of a new wild-type amine dehydrogenase used in an enzyme-catalyzed synthesis of an enantiomerically pure primary amine.

scholarly journals Economy Assessment for the Chiral Amine Production with Comparison of Reductive Amination and Transamination Routes by Multi-Enzyme System

Catalysts ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 1451
Author(s):  
Heyu Huo ◽  
Guangxiao Yao ◽  
Shizhen Wang
Keyword(s):  
Reductive Amination ◽  
Nadh Oxidase ◽  
Glucose Dehydrogenase ◽  
Economic Assessment ◽  
Building Blocks ◽  
Unit Price ◽  
Chiral Amine ◽  
Process Comparison ◽  
Amine Dehydrogenase

Chiral amines are key building blocks for pharmaceuticals. Economic assessment of commercial potential of bioprocesses is needed for guiding research. Biosynthesis of (S)-α-methylbenzylamine (MBA) was selected as case study. For transamination route, transaminase coupled with glucose dehydrogenase and lactate dehydrogenase catalyzed the reaction with NADH (Nicotinamide adenine dinucleotide) regeneration. Amine dehydrogenase coupled with NADH oxidase, which catalyzed the reductive amination process. Comparison of biosynthesis cost by reductive amination and transamination routes was carried out. Economic assessment based on the framework of cost analysis and preliminary process information revealed that cost is greatly dependent on enzyme price. The results indicated that enhancing the activity of amine dehydrogenase by 4–5 folds can drop the unit price of reductive amination to $0.5–0.6/g, which make it competitive with transamination route.

Reductive Amination of Biobased Levulinic Acid to Unnatural Chiral γ-Amino Acid Using an Engineered Amine Dehydrogenase

2020 ◽  
Vol 8 (46) ◽  
pp. 17054-17061
Author(s):  
Rui-Feng Cai ◽  
Lei Liu ◽  
Fei-Fei Chen ◽  
Aitao Li ◽  
Jian-He Xu ◽  
...  
Keyword(s):  
Amino Acid ◽  
Levulinic Acid ◽  
Reductive Amination ◽  
Amine Dehydrogenase

scholarly journals Biosynthesis of Chiral Amino Alcohols via an Engineered Amine Dehydrogenase in E. coli

2022 ◽  
Vol 9 ◽  
Author(s):  
Feifei Tong ◽  
Zongmin Qin ◽  
Hongyue Wang ◽  
Yingying Jiang ◽  
Junkuan Li ◽  
...  
Keyword(s):  
Reductive Amination ◽  
Asymmetric Reduction ◽  
Amino Alcohols ◽  
Saturation Mutagenesis ◽  
Preparative Scale ◽  
One Step ◽  
Amine Dehydrogenase ◽  
The One ◽  
Total Turnover ◽  
Chiral Amino Alcohols

Chiral amino alcohols are prevalent synthons in pharmaceuticals and synthetic bioactive compounds. The efficient synthesis of chiral amino alcohols using ammonia as the sole amino donor under mild conditions is highly desired and challenging in organic chemistry and biotechnology. Our previous work explored a panel of engineered amine dehydrogenases (AmDHs) derived from amino acid dehydrogenase (AADH), enabling the one-step synthesis of chiral amino alcohols via the asymmetric reductive amination of α-hydroxy ketones. Although the AmDH-directed asymmetric reduction is in a high stereoselective manner, the activity is yet fully excavated. Herein, an engineered AmDH derived from a leucine dehydrogenase from Sporosarcina psychrophila (SpAmDH) was recruited as the starting enzyme, and the combinatorial active-site saturation test/iterative saturation mutagenesis (CAST/ISM) strategy was applied to improve the activity. After three rounds of mutagenesis in an iterative fashion, the best variant wh84 was obtained and proved to be effective in the asymmetric reductive amination of 1-hydroxy-2-butanone with 4-fold improvements in kcat/Km and total turnover number (TTN) values compared to those of the starting enzyme, while maintaining high enantioselectivity (ee >99%) and thermostability (T5015 >53°C). In preparative-scale reaction, the conversion of 100 and 200 mM 1-hydroxy-2-butanone catalyzed by wh84 was up to 91–99%. Insights into the source of an enhanced activity were gained by the computational analysis. Our work expands the catalytic repertoire and toolbox of AmDHs.

One-pot reductive amination of carboxylic acids: a sustainable method for primary amine synthesis

2020 ◽  
Vol 22 (15) ◽  
pp. 5105-5114
Author(s):  
Robin Coeck ◽  
Dirk E. De Vos
Keyword(s):  
Carboxylic Acids ◽  
Primary Amine ◽  
Reductive Amination ◽  
One Pot ◽  
Tungsten Catalysts

Recyclable ruthenium–tungsten catalysts perform excellently for the direct reductive amination of carboxylic acids, with up to 96% yield.

Asymmetric Reductive Amination of Structurally Diverse Ketones with Ammonia Using a Spectrum-Extended Amine Dehydrogenase

ACS Catalysis ◽  
2021 ◽  
pp. 14274-14283
Author(s):  
Dong-Hao Wang ◽  
Qi Chen ◽  
Sai-Nan Yin ◽  
Xu-Wei Ding ◽  
Yu-Cong Zheng ◽  
...  
Keyword(s):  
Reductive Amination ◽  
Amine Dehydrogenase

Synthesis of enantiomerically pure 3-amino-1,2-diols by reductive amination of racemic 2,3-dialkoxyketones

1998 ◽  
Vol 39 (45) ◽  
pp. 8277-8280 ◽  
Author(s):  
Pierre Hutin ◽  
Yves Petit ◽  
Marc Larchevêque
Keyword(s):  
Reductive Amination ◽  
Enantiomerically Pure

scholarly journals Mycobacterium smegmatisl-Alanine Dehydrogenase (Ald) Is Required for Proficient Utilization of Alanine as a Sole Nitrogen Source and Sustained Anaerobic Growth

2002 ◽  
Vol 184 (18) ◽  
pp. 5001-5010 ◽  
Author(s):  
Zhengyu Feng ◽  
Nancy E. Cáceres ◽  
Gautam Sarath ◽  
Raúl G. Barletta
Keyword(s):  
Enzyme Activity ◽  
Nitrogen Source ◽  
Type Strain ◽  
Growth Stage ◽  
Reductive Amination ◽  
Wild Type Strain ◽  
Anaerobic Growth ◽  
Sole Nitrogen Source ◽  
Wild Type ◽  
Anaerobic Conditions

ABSTRACT NAD(H)-dependent l-alanine dehydrogenase (EC 1.4.1.1) (Ald) catalyzes the oxidative deamination of l-alanine and the reductive amination of pyruvate. To assess the physiological role of Ald in Mycobacterium smegmatis, we cloned the ald gene, identified its promoter, determined the protein expression levels, and analyzed the combined effects of nutrient supplementation, oxygen availability, and growth stage on enzyme activity. High Ald activities were observed in cells grown in the presence of l- or d-alanine regardless of the oxygen availability and growth stage. In exponentially growing cells under aerobic conditions, supplementation with alanine resulted in a 25- to 50-fold increase in the enzyme activity. In the absence of alanine supplementation, 23-fold-higher Ald activities were observed in cells grown exponentially under anaerobic conditions. Furthermore, M. smegmatis ald null mutants were constructed by targeted disruption and were shown to lack any detectable Ald activity. In contrast, the glycine dehydrogenase (EC 1.4.1.10) (Gdh) activity in mutant cells remained at wild-type levels, indicating that another enzyme protein is responsible for the physiologically relevant reductive amination of glyoxylate. The ald mutants grew poorly in minimal medium with l-alanine as the sole nitrogen source, reaching a saturation density 100-fold less than that of the wild-type strain. Likewise, mutants grew to a saturation density 10-fold less than that of the wild-type strain under anaerobic conditions. In summary, the phenotypes displayed by the M. smegmatis ald mutants suggest that Ald plays an important role in both alanine utilization and anaerobic growth.

scholarly journals Comparison of Family 9 Cellulases from Mesophilic and Thermophilic Bacteria

2010 ◽  
Vol 77 (4) ◽  
pp. 1436-1442 ◽  
Author(s):  
Florence Mingardon ◽  
John D. Bagert ◽  
Cyprien Maisonnier ◽  
Devin L. Trudeau ◽  
Frances H. Arnold
Keyword(s):  
High Temperatures ◽  
Catalytic Domain ◽  
Thermophilic Bacteria ◽  
Cellulose Degradation ◽  
Cellulase Activity ◽  
Crystalline Cellulose ◽  
Wild Type ◽  
Thermostable Enzymes ◽  
Highly Active ◽  
Homology Module

ABSTRACTCellulases containing a family 9 catalytic domain and a family 3c cellulose binding module (CBM3c) are important components of bacterial cellulolytic systems. We measured the temperature dependence of the activities of three homologs:Clostridium cellulolyticumCel9G,Thermobifida fuscaCel9A, andC. thermocellumCel9I. To directly compare their catalytic activities, we constructed six new versions of the enzymes in which the three GH9-CBM3c domains were fused to a dockerin both with and without aT. fuscafibronectin type 3 homology module (Fn3). We studied the activities of these enzymes on crystalline cellulose alone and in complex with a miniscaffoldin containing a cohesin and a CBM3a. The presence of Fn3 had no measurable effect on thermostability or cellulase activity. The GH9-CBM3c domains of Cel9A and Cel9I, however, were more active than the wild type when fused to a dockerin complexed to scaffoldin. The three cellulases in complex have similar activities on crystalline cellulose up to 60°C, butC. thermocellumCel9I, the most thermostable of the three, remains highly active up to 80°C, where its activity is 1.9 times higher than at 60°C. We also compared the temperature-dependent activities of different versions of Cel9I (wild type or in complex with a miniscaffoldin) and found that the thermostable CBM is necessary for activity on crystalline cellulose at high temperatures. These results illustrate the significant benefits of working with thermostable enzymes at high temperatures, as well as the importance of retaining the stability of all modules involved in cellulose degradation.

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