ABSTRACT
Early trials have demonstrated great potential for the use of duckweed (family Lemnaceae) as the next generation of energy plants for the production of biofuels. Achieving this technological advance demands research to develop novel bioengineering microorganisms that can ferment duckweed feedstock to produce higher alcohols. In this study, we used relevant genes to transfer five metabolic pathways of isoleucine, leucine and valine from the yeast Saccharomyces cerevisiae into the bioengineered microorganism Corynebacterium crenatum. Experimental results showed that the bioengineered strain was able to produce 1026.61 mg/L of 2-methyl-1-butanol by fermenting glucose, compared to 981.79 mg/L from the acid hydrolysates of duckweed. The highest isobutanol yields achieved were 1264.63 mg/L from glucose and 1154.83 mg/L from duckweed, and the corresponding highest yields of 3-methyl-1-butanol were 748.35 and 684.79 mg/L. Our findings demonstrate the feasibility of using bioengineered C. crenatum as a platform to construct a bacterial strain that is capable of producing higher alcohols. We have also shown the promise of using duckweed as the basis for developing higher alcohols, illustrating that this group of plants represents an ideal fermentation substrate that can be considered the next generation of alternative energy feedstocks.
Subject(s)
Alcohols/metabolism , Araceae/chemistry , Biofuels , Corynebacterium/metabolism , Metabolic Engineering/methods , Plant Extracts/metabolism , Butanols/metabolism , Corynebacterium/genetics , Fermentation , Gene Expression , Glucose/metabolism , Hydrolysis , Isoleucine/metabolism , Leucine/metabolism , Metabolic Networks and Pathways/genetics , Reproducibility of Results , Reverse Transcriptase Polymerase Chain Reaction , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/genetics , Saccharomyces cerevisiae Proteins/metabolism , Starch/metabolism , Valine/metabolismABSTRACT
The effect of temperature, light intensity, nitrogen and phosphorus concentrations on the biomass and starch content of duckweed (Landoltia punctata OT, Lemna minor OT) in monoculture and mixture were assessed. Low light intensity promoted more starch accumulation in mixture than in monoculture. The duckweed in mixture had higher biomass and nutrient removal efficiency than those in monoculture in swine wastewater. Moreover, the ability of L. punctata C3, L. minor C2, Spirodela polyrhiza C1 and their mixtures to recovery nutrients and their biomass were analyzed. Results showed that L. minor C2 had the highest N and P content, while L. punctata C3 had the highest starch content, and the mixture of L. punctata C3 and L. minor C2 had the greatest nutrient removal rate and the highest biomass. Compared with L. punctata C3 and L. minor C2 in monoculture, their biomass in mixture increased by 17.0% and 39.8%, respectively.