NADH/NADPH bi-cofactor-utilizing and thermoactive ketol-acid reductoisomerase from Sulfolobus acidocaldarius

Chen, Chin‐Yu; Ko, Tzu‐Ping; Lin, Kuan-Fu; Lin, Bo‐Lin; Huang, Chun‐Hsiang; Chiang, Cheng-Hung; Horng, Jia‐Cherng

doi:10.1038/s41598-018-25361-4

Cited by 12 publications

(20 citation statements)

References 47 publications

(113 reference statements)

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“…The mechanism of how BCAA concentration is increased by Halanaerobium can be explained by the enzymatic character of ketol-acid reductoisomerase (KARI). KARI is a key enzyme in the Ile and Leu/Val biosynthetic pathways ( Chen et al, 2018 ) and is known to use NADPH as a coenzyme ( Fig. S2 ).…”

Section: Discussionmentioning

confidence: 99%

The effects of vegetable pickling conditions on the dynamics of microbiota and metabolites

Sawada¹,

Koyano

Yamamoto

et al. 2021

PeerJ

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Background Salting is a traditional procedure for producing pickled vegetables. Salting can be used as a pretreatment, for safe lactic acid fermentation and for salt stock preparation. This study aimed to provide valuable knowledge to improve pickle production by investigating the dynamics of microbiota and metabolites during the pretreatment and salt stock preparation processes, which have previously been overlooked. The differences in these process conditions would be expected to change the microbiota and consequently influence the content of metabolites in pickles. Methods Samples, collected from eight commercial pickle manufacturers in Japan, consisted of the initial raw materials, pickled vegetables and used brine. The microbiota were analyzed by 16S rRNA sequencing and the metabolites quantified by liquid chromatograph-mass spectrometry. Statistical analyses helped to identify any significant differences between samples from the initial raw materials, pretreatment process and salt stock preparation process groups. Results Under pretreatment conditions, aerobic and facultative anaerobic bacteria were predominant, including Vibrio, a potentially undesirable genus for pickle production. Under salt stock preparation conditions, the presence of halophilic bacteria, Halanaerobium, suggested their involvement in the increase in pyruvate derivatives such as branched-chain amino acids (BCAA). PICRUSt analysis indicated that the enhanced production of BCAA in salt stock was caused not by quantitative but by qualitative differences in the biosynthetic pathway of BCAA in the microbiota. Conclusion The differences in the microbiota between pretreatment and previously studied lactic acid fermentation processes emphasized the importance of anaerobic conditions and low pH under moderate salinity conditions for assuring safe pickle production. The results from the salt stock preparation process suggested that the Halanaerobium present may provide a key enzyme in the BCAA biosynthetic pathway which prefers NADH as a coenzyme. This feature can enhance BCAA production under anaerobic conditions where NADH is in excess. The effects shown in this study will be important for adjusting pickling conditions by changing the abundance of bacteria to improve the quality of pickled vegetables.

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Section: Discussionmentioning

confidence: 99%

The effects of vegetable pickling conditions on the dynamics of microbiota and metabolites

Sawada¹,

Koyano

Yamamoto

et al. 2021

PeerJ

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“…KARIs are structurally composed of an N-terminal Rossmann domain and a Cterminal "knot" domain [1,7]. They can be distinguished in two classes: the short class I gathering dimeric and dodecameric enzymes, and the monomeric or tetrameric longer class II KARIs [1,[6][7][8][9][10][11][12][13][14]. Prokaryotes can harbour class I or II enzymes, while eukaryotes only contain class II.…”

Section: Introductionmentioning

confidence: 99%

Structural Rearrangements of a Dodecameric Ketol-Acid Reductoisomerase Isolated from a Marine Thermophilic Methanogen

et al. 2021

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Ketol-acid reductoisomerase (KARI) orchestrates the biosynthesis of branched-chain amino acids, an elementary reaction in prototrophic organisms as well as a valuable process in biotechnology. Bacterial KARIs belonging to class I organise as dimers or dodecamers and were intensively studied to understand their remarkable specificity towards NADH or NADPH, but also to develop antibiotics. Here, we present the first structural study on a KARI natively isolated from a methanogenic archaea. The dodecameric structure of 0.44-MDa was obtained in two different conformations, an open and close state refined to a resolution of 2.2-Å and 2.1-Å, respectively. These structures illustrate the conformational movement required for substrate and coenzyme binding. While the close state presents the complete NADP bound in front of a partially occupied Mg2+-site, the Mg2+-free open state contains a tartrate at the nicotinamide location and a bound NADP with the adenine-nicotinamide protruding out of the active site. Structural comparisons show a very high conservation of the active site environment and detailed analyses point towards few specific residues required for the dodecamerisation. These residues are not conserved in other dodecameric KARIs that stabilise their trimeric interface differently, suggesting that dodecamerisation, the cellular role of which is still unknown, might have occurred several times in the evolution of KARIs.

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“…13 CRs have been successfully applied industrially for the synthesis of various chiral alcohols. However, the industrial potential of CRs was largely restricted by the cofactor regeneration efficiency and the expensive price of the cofactor NAD(P)H. [14][15][16] In situ cofactor regeneration is considered essential for the economic viability of industrial scale biotransformation. Kizaki In our previous study, a recombinant carbonyl reductase CR125 was obtained from Lactobacillus kefiri to catalyze 150 g/L ET-4 to (S)-ET-5 with the conversion of 99.1% and the e.e.…”

Section: Introductionmentioning

confidence: 99%

“…CRs have been successfully applied industrially for the synthesis of various chiral alcohols. However, the industrial potential of CRs was largely restricted by the cofactor regeneration efficiency and the expensive price of the cofactor NAD(P)H 14‐16 . In situ cofactor regeneration is considered essential for the economic viability of industrial scale biotransformation.…”

Section: Introductionmentioning

confidence: 99%

Efficient production of an ezetimibe intermediate using carbonyl reductase coupled with glucose dehydrogenase

Zhang

Zhou

et al. 2020

Biotechnology Progress

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Ezetimibe is a top-selling hypolipidemic drug for the treatment of cardiovascular diseases. Biosynthesis of (4S)-3-[(5S)-5-(4-fluorophenyl)-5-hydroxypentanoyl]-4-phenyl-1,3-oxazolidin-2-one ((S)-ET-5) using carbonyl reductase has shown advantages including high catalytic efficiency, excellent stereoselectivity, mild reaction conditions, and environmental friendness, and was considered as the key step for ezetimibe production. The regeneration efficiency of the cofactor, nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) is one of the main restricted factor. Recombinant Escherichia coli strain (smCR125) coexpressing carbonyl reductase (CR125) and glucose dehydrogenase were successfully constructed and applied for the production of (S)-ET-5 for the first time. Without extra addition of the coenzyme NADPH, the yield of 99.8% and the enantiomeric excess (e.e.) of 99.9% were achieved under ET-4 concentration of 200 g/L. Using a substrate fed-batch strategy, under the optimal conditions, the substrate ET-4 concentration was increased to 250 g/L with the yield of 98.9% and the e.e. of 99.9% after 12 hr reaction. The space-time yield of 494.5 g L −1 d −1 and the space-time yield per gram biocatalyst of 24.7 g L −1 d −1 g −1 DCW were achieved, which were higher than ever reported for the biosynthesis of the ezetimibe intermediate.

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NADH/NADPH bi-cofactor-utilizing and thermoactive ketol-acid reductoisomerase from Sulfolobus acidocaldarius

Cited by 12 publications

References 47 publications

The effects of vegetable pickling conditions on the dynamics of microbiota and metabolites

The effects of vegetable pickling conditions on the dynamics of microbiota and metabolites

Structural Rearrangements of a Dodecameric Ketol-Acid Reductoisomerase Isolated from a Marine Thermophilic Methanogen

Efficient production of an ezetimibe intermediate using carbonyl reductase coupled with glucose dehydrogenase

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