Metabolic engineering of Zymomonas mobilis for 2,3-butanediol production from lignocellulosic biomass sugars

Yang, Shihui; Mohagheghi, Ali; Franden, Mary Ann; Chou, Yat Chen.; Chen, Xiaowen; Dowe, Nancy; Himmel, Michael E.; Zhang, Min

doi:10.1186/s13068-016-0606-y

Cited by 130 publications

(146 citation statements)

References 90 publications

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“…(g L −1 )Pro. b (g L −1 h −1 )Refs. B. subtilis IPE5-4-UD-4Pretreated corncobBatch SSF, 5-L bioreactor50; 5022.760.38This study K. pneumoniae CICC 10011Pretreated Jerusalem artichoke stalksBatch SSF, 5-L bioreactor37; 3713.470.20[9] E. coli CelluloseBatch SSF, shake flask37; 372.700.02[13] E. cloacae SDM 53Pretreated corn stoverFed-batch SHF, 7.5-L bioreactorNA; 3745.601.52[23] K. pneumoniae CICC 10011Pretreated Jerusalem artichoke stalk and tuberFed-batch SHF, 5-L bioreactor50–55; 3711.400.16[9] E. coli DSM02-BPretreated brown seaweedFed-batch SHF, shake flask50; 374.800.07[12] E. coli mlc-XT7-LAFC-YSDPretreated Japanese cedar wood chipsBatch SHF, shake flask37; 3019.000.16[24] K. pneumoniae CICC 10011Pretreated Jerusalem artichoke stalk and tuberBatch SHF, 5-L bioreactor50–55; 3711.800.23[9]Engineered Zymomonas mobilis 22C–BC5Pretreated corn stoverBatch SHF, shake flask50; 33> 10.00NA[10] Tem...…”

Section: Discussionmentioning

confidence: 99%

Conversion of cellulose and hemicellulose of biomass simultaneously to acetoin by thermophilic simultaneous saccharification and fermentation

Jia

Peng

Liu

et al. 2017

Biotechnol Biofuels

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BackgroundAcetoin (3-hydroxy-2-butanone), the precursor of biofuel 2,3-butanediol, is an important bio-based platform chemical with wide applications. Fermenting the low-cost and renewable plant biomass is undoubtedly a promising strategy for acetoin production. Isothermal simultaneous saccharification and fermentation (SSF) is regarded as an efficient method for bioconversion of lignocellulosic biomass, in which the temperature optima fitting for both lignocellulose-degrading enzymes and microbial strains.ResultsA thermotolerant (up to 52 °C) acetoin producer Bacillus subtilis IPE5-4 which simultaneously consumed glucose and xylose was isolated and identified. By compound mutagenesis, the mutant IPE5-4-UD-4 with higher acetoin productivity was selected. When fermenting at 50 °C in a 5-L bioreactor using glucose as the feedstock by strain IPE5-4-UD-4, the acetoin concentration reached 28.83 ± 0.91 g L−1 with the acetoin yield and productivity of 0.34 g g−1 glucose and 0.60 g L−1 h−1, respectively. Furthermore, an optimized and thermophilic SSF process operating at 50 °C was conducted for acetoin production from alkali-pretreated corncob (APC). An acetoin concentration of 12.55 ± 0.28 g L−1 was achieved by strain IPE5-4-UD-4 in shake flask SSF, with the acetoin yield and productivity of 0.25 g g−1 APC and 0.17 g L−1 h−1. Meanwhile, the utilization of cellulose and hemicellulose in the SSF approach reached 96.34 and 93.29%, respectively. When further fermented at 50 °C in a 5-L bioreactor, the concentration of acetoin reached the maximum of 22.76 ± 1.16 g L−1, with the acetoin yield and productivity reaching, respectively, 0.46 g g−1 APC and 0.38 g L−1 h−1. This was by far the highest acetoin yield in SSF from lignocellulosic biomass.ConclusionsThis thermophilic SSF process provided an efficient and economical route for acetoin production from lignocellulosic biomass at ideal temperature for both enzymatic hydrolysis and microbial fermentation.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-017-0924-8) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Conversion of cellulose and hemicellulose of biomass simultaneously to acetoin by thermophilic simultaneous saccharification and fermentation

Jia

Peng

Liu

et al. 2017

Biotechnol Biofuels

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“…Ethanol production from lignocellulosic biomass provides opportunities for exploring the advantage of Z. mibilis , because only glucose is released from cellulose hydrolysis, and in the meantime this species can be engineered with xylose isomerase to convert xylose to xylulose directly without cofactor imbalance and xylitol accumulation that are intrinsic to S. cerevisiae engineered with xylose reductase and xylitol dehydrogenase for ethanol production from xylose (Jeffries, ; M. Zhang, Eddy, Deanda, Finkelstein, & Picataggio, ). In addition, Z. mobilis is also a potential host to be engineered for biorefinery to produce biobased chemicals such as 2, 3‐butanediol (M. He et al, ; Yang, Fei et al, ; Yang, Mohagheghi et al ).…”

Section: Introductionmentioning

confidence: 99%

Contribution of cellulose synthesis, formation of fibrils and their entanglement to the self‐flocculation of Zymomonas mobilis

Xia

Liu

Zhao

et al. 2018

Biotech & Bioengineering

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Due to the unique Entner-Doudoroff pathway, Zymomonas mobilis has been acknowledged as a potential host to be engineered for biorefinery to produce biofuels and biobased chemicals. The self-flocculation of Z. mobilis can make the bacterial cells self-immobilized within bioreactors for high density to improve product productivities, and in the meantime enhance their tolerance to stresses, particularly product inhibition and the toxicity of byproducts released during the pretreatment of lignocellulosic biomass. In this work, we explored mechanism underlying such a phenotype with the self-flocculating strain ZM401 developed from the regular non-flocculating strain ZM4. Cellulase de-flocculation and the restoration of the self-flocculating phenotype for the de-flocculated bacterial cells subjected to culture confirmed the essential role of cellulose biosynthesis in the self-flocculation of ZM401. Furthermore, the deactivation of both Type I and Type IV restriction-modification systems was performed for ZM4 and ZM401 to improve their transformation efficiencies. Comparative genome analysis detected the deletion of a thymine from ZMO1082 in ZM401, leading to a frame-shift mutation for the putative gene to be integrated into the neighboring downstream gene ZMO1083 encoding the catalytic subunit A of cellulose synthase, and consequently created a new gene to encode a larger transmembrane protein BcsA_401 for more efficient synthesis of cellulose as well as the development of cellulose fibrils and their entanglement for the self-flocculation of the mutant. These speculations were confirmed by the morphological observation of the bacterial cells under scanning electron microscopy, the impact of the gene deletion on the self-flocculation of ZM401, and the restoration of the self-flocculating phenotype of ZM401 ΔbcsA by the gene complementation. The progress will lay a foundation not only for fundamental research in deciphering molecular mechanisms underlying the self-flocculation of Z. mobilis and stress tolerance associated with the morphological change but also for technological innovations in engineering non-flocculating Z. mobilis and other bacterial species with the self-flocculating phenotype.

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“…However, several problems are associated with the constitutive promoters for metabolic engineering practices. For example, the constant expression of heterologous genes would accumulate intermediate metabolites which may be toxic to cells , and strong constitutive promoters could cause the energy and metabolic burdens and therefore impact the cellular growth .…”

Section: Introductionmentioning

confidence: 99%

“…Up to now, a few inducible promoters are tested and used in Z. mobilis . Only one inducible promoter, tetracycline‐inducible promoter P tet , was successfully utilized, which was used to direct metabolic flux to produce 2,3‐butanediol and to construct the inducible dual reporter–gene system .…”

Section: Introductionmentioning

confidence: 99%

Identification and characterization of ethanol‐inducible promoters of Zymomonas mobilis based on omics data and dual reporter–gene system

Yang

Rong

Song

et al. 2019

Biotech and App Biochem

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Zymomonas mobilis is a model bacterial ethanologen and has been engineered to produce lignocellulosic biofuels and biochemicals such as 2,3‐butanediol. We have previously identified promoters of different strengths using systems biology datasets and characterized them using the flow cytometry–based dual reporter–gene system. Here, we further demonstrated the capability of applying the dual reporter–gene system and omics datasets on discovering inducible promoters. Ten candidate ethanol‐inducible promoters were identified through omics datasets mining and clustering. Using the dual reporter–gene system, these promoters were characterized under natural growth, ethanol stress, and ethanol‐induced condition to investigate the transcriptional strength and ethanol inducibility. The results demonstrated that three promoters of P0405, P0435, and P0038 driving the expression of native genes of ZMO0405, ZMO0435, and ZMO0038, correspondingly, are potential ethanol‐responsive promoters and may be growth related. This study not only identified and verified three ethanol‐inducible promoters as biological parts, which can be used to synchronize the expression of heterologous pathway genes with the ethanol production process of Z. mobilis, but also demonstrated the power of combining omics datasets and dual reporter–gene system to identify biological parts for metabolic engineering and synthetic biology applications in Z. mobilis and related microorganisms.

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Metabolic engineering of Zymomonas mobilis for 2,3-butanediol production from lignocellulosic biomass sugars

Cited by 130 publications

References 90 publications

Conversion of cellulose and hemicellulose of biomass simultaneously to acetoin by thermophilic simultaneous saccharification and fermentation

Conversion of cellulose and hemicellulose of biomass simultaneously to acetoin by thermophilic simultaneous saccharification and fermentation

Contribution of cellulose synthesis, formation of fibrils and their entanglement to the self‐flocculation of Zymomonas mobilis

Identification and characterization of ethanol‐inducible promoters of Zymomonas mobilis based on omics data and dual reporter–gene system

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