Coenzyme A-acylating propionaldehyde dehydrogenase (PduP) from Lactobacillus reuteri: Kinetic characterization and molecular modeling

Sabet-Azad, Ramin; Linares-Pastén, Javier A.; Torkelson, Lisa; Sardari, Roya R.R.; Hatti‐Kaul, Rajni

doi:10.1016/j.enzmictec.2013.05.007

Cited by 21 publications

(21 citation statements)

References 32 publications

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“…3HPA besides being toxic to the cells is also inhibitory to the PduP enzyme at a concentration exceeding 0.6 g/L [34]. Moreover, high glycerol and 1,3PDO concentrations inhibit glycerol dehydratase activity [49].…”

Section: Resultsmentioning

confidence: 99%

“…After 1 h of batch biotransformation, fed-batch mode was started by feeding glycerol (50 g/L) at a rate of 12 mL/h (0.6 g gly /h) for 10 h. Subsequently, the feeding rate was increased to 31.1 mL/h (1.6 g gly /h) for 10 h, and finally to 50 mL/h (2.5 g gly /h) for 10 h. The biotransformation was performed at 37˚C, pH 7, 500 rpm, with continuous nitrogen bubbling to maintain anaerobic conditions. The pH was chosen based on the reported optimum for some of the enzymes of the Pdu pathway [34,50]. …”

Section: Methodsmentioning

confidence: 99%

“…The production of 3HP alone or in a mixture with 1,3PDO as main products using a wild-type L. reuteri has however not been successful [22,33], mainly due to the accumulation to toxic levels of the intermediate 3HPA [22,28] and its inhibitory effect on one of the enzymes of the oxidative pathway [34], as well as the diversion of glycerol to dihydroxyacetone (DHA) catalyzed by a glycerol dehydrogenase [33]. The co-production of 3HP and 1,3PDO was only possible after the gene encoding the enzyme glycerol dehydrogenase was knocked out [22].…”

Section: Introductionmentioning

confidence: 99%

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Flux analysis of the Lactobacillus reuteri propanediol-utilization pathway for production of 3-hydroxypropionaldehyde, 3-hydroxypropionic acid and 1,3-propanediol from glycerol

et al. 2014

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BackgroundLactobacillus reuteri converts glycerol to 3-hydroxypropionic acid (3HP) and 1,3-propanediol (1,3PDO) via 3-hydroxypropionaldehyde (3HPA) as an intermediate using enzymes encoded in its propanediol-utilization (pdu) operon. Since 3HP, 1,3PDO and 3HPA are important building blocks for the bio-based chemical industry, L. reuteri can be an attractive candidate for their production. However, little is known about the kinetics of glycerol utilization in the Pdu pathway in L. reuteri. In this study, the metabolic fluxes through the Pdu pathway were determined as a first step towards optimizing the production of 3HPA, and co-production of 3HP and 1,3PDO from glycerol. Resting cells of wild-type (DSM 20016) and recombinant (RPRB3007, with overexpressed pdu operon) strains were used as biocatalysts.ResultsThe conversion rate of glycerol to 3HPA by the resting cells of L. reuteri was evaluated by in situ complexation of the aldehyde with carbohydrazide to avoid the aldehyde-mediated inactivation of glycerol dehydratase. Under operational conditions, the specific 3HPA production rate of the RPRB3007 strain was 1.9 times higher than that of the wild-type strain (1718.2 versus 889.0 mg/gCDW.h, respectively). Flux analysis of glycerol conversion to 1,3PDO and 3HP in the cells using multi-step variable-volume fed-batch operation showed that the maximum specific production rates of 3HP and 1,3PDO were 110.8 and 93.7 mg/gCDW.h, respectively, for the wild-type strain, and 179.2 and 151.4 mg/gCDW.h, respectively, for the RPRB3007 strain. The cumulative molar yield of the two compounds was ~1 mol/mol glycerol and their molar ratio was ~1 mol3HP/mol1,3PDO. A balance of redox equivalents between the glycerol oxidative and reductive pathway branches led to equimolar amounts of the two products.ConclusionsMetabolic flux analysis was a useful approach for finding conditions for maximal conversion of glycerol to 3HPA, 3HP and 1,3PDO. Improved specific production rates were obtained with resting cells of the engineered RPRB3007 strain, highlighting the potential of metabolic engineering to render an industrially sound strain. This is the first report on the production of 3HP and 1,3PDO as sole products using the wild-type or mutant L. reuteri strains, and has laid ground for further work on improving the productivity of the biotransformation process using resting cells.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Flux analysis of the Lactobacillus reuteri propanediol-utilization pathway for production of 3-hydroxypropionaldehyde, 3-hydroxypropionic acid and 1,3-propanediol from glycerol

et al. 2014

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“…Then a phosphotransacylase (PduL) converts 3-HP-CoA into 3-HPphosphate and simultaneously releases the Coenzyme A. Actually, a propionate kinase (PduW) leads to 3-HP and ATP [13,15,25]. 3-HPA can also be reduced in 1,3-propanediol (1,3-PDO) by a 1,3-PDO oxidoreductase (PduQ) requiring NADH, H + [26] (Fig.…”

Section: Introductionmentioning

confidence: 97%

“…1), the ratio between 3-HP and 1,3-PDO depending in part on the NADH, H + availability. The whole enzymes involved in this glycerol metabolic pathway are encoded in the propanediol utilization (pdu) operon [25,27]. Among the products of this metabolic pathway, only the toxicity of 3-HPA towards the producing strains has been studied in details [28][29][30][31].…”

Section: Introductionmentioning

confidence: 99%

Diversity of Lactobacillus reuteri Strains in Converting Glycerol into 3-Hydroxypropionic Acid

Burgé¹,

Saulou‐Bérion²,

Moussa³

et al. 2015

Appl Biochem Biotechnol

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The present study aims at comparing the performances of three Lactobacillus reuteri strains (DSM 20016, DSM 17938, and ATCC 53608) in producing 3-hydroxypropionic acid (3-HP) from glycerol and at exploring inhibition phenomena during this bioconversion. Differences were highlighted between the three strains in terms of 3-HP production yield, kinetics of substrate consumption, and metabolite production. With a maximal productivity in non-optimal conditions (free pH) around 2 g.L(-1).h(-1) of 3-HP and 4 g.L(-1).h(-1) of 3-hydroxypropionaldehyde (3-HPA) depending on the strain, this study confirmed the potential of L. reuteri for the biotechnological production of 3-HP. Moreover, the molar ratios of 3-HP to 1,3-propanediol (1,3-PDO) obtained for the three strains (comprised between 1.25 and 1.65) showed systematically a higher 3-HP production. From these results, the DSM 17938 strain appeared to be the most promising strain. The impact of glycerol bioconversion on the bacteria's physiological state (a decrease of around 40 % in DSM 17938 cells showing an enzymatic activity after 3 h) and survival (total loss of cultivability after 2 or 3 h depending on the strains) was revealed and discussed. The effect of each metabolite on L. reuteri DSM 17938 was further investigated, displaying a drastic inhibition caused by 3-HPA, while 3-HP induced lower impact and only at acidic pH.

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Recent advances, challenges and metabolic engineering strategies in the biosynthesis of 3‐hydroxypropionic acid

Liang

Sun

Nie

et al. 2022

Biotech & Bioengineering

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As an attractive and valuable platform chemical, 3‐hydroxypropionic acid (3‐HP) can be used to produce a variety of industrially important commodity chemicals and biodegradable polymers. Moreover, the biosynthesis of 3‐HP has drawn much attention in recent years due to its sustainability and environmental friendliness. Here, we focus on recent advances, challenges, and metabolic engineering strategies in the biosynthesis of 3‐HP. While glucose and glycerol are major carbon sources for its production of 3‐HP via microbial fermentation, other carbon sources have also been explored. To increase yield and titer, synthetic biology and metabolic engineering strategies have been explored, including modifying pathway enzymes, eliminating flux blockages due to byproduct synthesis, eliminating toxic byproducts, and optimizing via genome‐scale models. This review also provides insights on future directions for 3‐HP biosynthesis.

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Coenzyme A-acylating propionaldehyde dehydrogenase (PduP) from Lactobacillus reuteri: Kinetic characterization and molecular modeling

Cited by 21 publications

References 32 publications

Flux analysis of the Lactobacillus reuteri propanediol-utilization pathway for production of 3-hydroxypropionaldehyde, 3-hydroxypropionic acid and 1,3-propanediol from glycerol

Flux analysis of the Lactobacillus reuteri propanediol-utilization pathway for production of 3-hydroxypropionaldehyde, 3-hydroxypropionic acid and 1,3-propanediol from glycerol

Diversity of Lactobacillus reuteri Strains in Converting Glycerol into 3-Hydroxypropionic Acid

Recent advances, challenges and metabolic engineering strategies in the biosynthesis of 3‐hydroxypropionic acid

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