Construction of engineered RuBisCO Kluyveromyces marxianus for a dual microbial bioethanol production system

Ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) genes play important roles in CO2 fixation and redox balancing in photosynthetic bacteria. In the present study, the kefir yeast Kluyveromyces marxianus 4G5 was used as host for the transformation of form I and form II RubisCO genes derived f...

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Bibliographic Details
Published in:PloS one 2021-03, Vol.16 (3), p.e0247135-e0247135
Main Authors: Ha-Tran, Dung Minh, Lai, Rou-Yin, Nguyen, Trinh Thi My, Huang, Eugene, Lo, Shou-Chen, Huang, Chieh-Chen
Format: Article
Language:English
Subjects:
Alcohol
Alcohol, Denatured
Biofuels
Biology and Life Sciences
Biomass
Broths
Carbon dioxide
Carbon dioxide fixation
Cell culture
Chemical properties
Cloning
Construction
Crop yield
Enzymatic activity
Enzymes
Ethanol
Genes
Genetic aspects
Genomes
Glucose
High temperature
Homologous recombination
Homology
Life sciences
Lignocellulose
Microorganisms
Physical Sciences
Plant biomass
Plasmids
Research and Analysis Methods
Ribulose-bisphosphate carboxylase
Rubisco
Transcription
Yeast
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Summary:Ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) genes play important roles in CO2 fixation and redox balancing in photosynthetic bacteria. In the present study, the kefir yeast Kluyveromyces marxianus 4G5 was used as host for the transformation of form I and form II RubisCO genes derived from the nonsulfur purple bacterium Rhodopseudomonas palustris using the Promoter-based Gene Assembly and Simultaneous Overexpression (PGASO) method. Hungateiclostridium thermocellum ATCC 27405, a well-known bacterium for its efficient solubilization of recalcitrant lignocellulosic biomass, was used to degrade Napier grass and rice straw to generate soluble fermentable sugars. The resultant Napier grass and rice straw broths were used as growth media for the engineered K. marxianus. In the dual microbial system, H. thermocellum degraded the biomass feedstock to produce both C5 and C6 sugars. As the bacterium only used hexose sugars, the remaining pentose sugars could be metabolized by K. marxianus to produce ethanol. The transformant RubisCO K. marxianus strains grew well in hydrolyzed Napier grass and rice straw broths and produced bioethanol more efficiently than the wild type. Therefore, these engineered K. marxianus strains could be used with H. thermocellum in a bacterium-yeast coculture system for ethanol production directly from biomass feedstocks.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0247135