Journal ArticleDOI
Enhanced cell-surface display and secretory production of cellulolytic enzymes with Saccharomyces cerevisiae Sed1 signal peptide.
TLDR
The utilization of the novel SP sequence derived from the Saccharomyces cerevisiae SED1 gene is a promising option for highly efficient cell‐surface display and secretory production of heterologous proteins in various yeast species.Abstract:
Recombinant yeast strains displaying aheterologous cellulolytic enzymes on their cell surfaces using a glycosylphosphatidylinositol (GPI) anchoring system are a promising strategy for bioethanol production from lignocellulosic materials. A crucial step for cell wall localization of the enzymes is the intracellular transport of proteins in yeast cells. Therefore, the addition of a highly efficient secretion signal sequence is important to increase the amount of the enzymes on the yeast cell surface. In this study, we demonstrated the effectiveness of a novel signal peptide (SP) sequence derived from the Saccharomyces cerevisiae SED1 gene for cell-surface display and secretory production of cellulolytic enzymes. Gene cassettes with SP sequences derived from S. cerevisiae SED1 (SED1SP), Rhizopus oryzae glucoamylase (GLUASP), and S. cerevisiae α-mating pheromone (MFα1SP) were constructed for cell-surface display of Aspergillus aculeatus β-glucosidase (BGL1) and Trichoderma reesei endoglucanase II (EGII). These gene cassettes were integrated into the S. cerevisiae genome. The recombinant strains with the SED1SP showed higher cell-surface BGL and EG activities than those with the conventional SP sequences (GLUASP and MFα1SP). The novel SP sequence also improved the secretory production of BGL and EG in S. cerevisiae. The extracellular BGL activity of the recombinant strains with the SED1SP was 1.3- and 1.9-fold higher than the GLUASP and MFα1SP strains, respectively. Moreover, the utilization of SED1SP successfully enhanced the secretory production of BGL in Pichia pastoris. The utilization of the novel SP sequence is a promising option for highly efficient cell-surface display and secretory production of heterologous proteins in various yeast species. Biotechnol. Bioeng. 2016;113: 2358-2366. © 2016 Wiley Periodicals, Inc.read more
Citations
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Journal ArticleDOI
Technological advances and applications of hydrolytic enzymes for valorization of lignocellulosic biomass.
Manisha,Sudesh Yadav +1 more
TL;DR: This article provides recent advancement towards the isolation and use of microbes for lignocellulosic biomass utilisation, microbes producing the hydrolytic enzymes, the modern age technologies used to manipulate and enhance the Hydrolytic enzyme activity and the applications of such enzymes in value added products development from lignOcellulOSic biomass.
Journal ArticleDOI
Large-scale production of enzymes for biotechnology uses.
TL;DR: The existing and emerging production approaches, applications, developments, and global need for enzymes are discussed, with special emphasis given to the predominantly utilized hydrolytic microbial enzymes in industrial bioprocesses.
Journal ArticleDOI
Enzyme mediated multi-product process: A concept of bio-based refinery
Bikash Kumar,Pradeep Verma +1 more
TL;DR: The role of different hydrolytic enzymes is important to the biobased refinery as mentioned in this paper, and the application of the advanced biotechnological approach can help in the development of a consolidated bioprocessing approach.
Journal ArticleDOI
Engineering microbes for direct fermentation of cellulose to bioethanol.
TL;DR: The recent advances in CBP microbes are reviewed and the efforts in strain improvement employing genetic engineering are focused on, including Saccharomyces cerevisiae and several other yeasts modified to express recombinant cellulases in media or display them on the cell surface for CBP of cellulose.
Journal ArticleDOI
Co-fermentation using recombinant Saccharomyces cerevisiae yeast strains hyper-secreting different cellulases for the production of cellulosic bioethanol
Cho-Ryong Lee,Cho-Ryong Lee,Bong Hyun Sung,Bong Hyun Sung,Kwang Mook Lim,Mi Jin Kim,Min Jeong Sohn,Jung Hoon Bae,Jung Hoon Sohn,Jung Hoon Sohn +9 more
TL;DR: The synergistic effect of the mixed culture of the four strains expressing the essential cellulases with the insoluble substrate Avicel and several types of cellulosic biomass was demonstrated to be effective and will contribute to the cost-effective production of bioenergy such as bioethanol and biochemicals from cellulose hydrolysis.
References
More filters
Journal ArticleDOI
Enzymatic assembly of DNA molecules up to several hundred kilobases
Daniel G. Gibson,Lei Young,Ray-Yuan Chuang,J. Craig Venter,Clyde A. Hutchison,Hamilton O. Smith +5 more
TL;DR: An isothermal, single-reaction method for assembling multiple overlapping DNA molecules by the concerted action of a 5′ exonuclease, a DNA polymerase and a DNA ligase is described.
Journal ArticleDOI
WoLF PSORT: protein localization predictor.
Paul Horton,Keun Joon Park,Takeshi Obayashi,Naoya Fujita,Hajime Harada,C. J. Adams-Collier,Kenta Nakai +6 more
TL;DR: WoLF PSORT converts protein amino acid sequences into numerical localization features; based on sorting signals, amino acid composition and functional motifs such as DNA-binding motifs, which allows a user to understand the evidence (or lack thereof) behind the predictions made for particular proteins.
Journal ArticleDOI
Protein expression in Pichia pastoris: recent achievements and perspectives for heterologous protein production.
TL;DR: This review refers to established tools in protein expression in P. pastoris and highlights novel developments in the areas of expression vector design, host strain engineering and screening for high-level expression strains.
Journal ArticleDOI
Fungal bioconversion of lignocellulosic residues; opportunities & perspectives.
TL;DR: The current status of the technology for bioconversion of biomass by fungi is reviewed, with focus on mutagenesis, co-culturing and heterologous gene expression attempts to improve fungal lignocellulolytic activities to create robust fungal strains.
Journal ArticleDOI
One-step transformation of yeast in stationary phase.
TL;DR: A fast yeast-transformation technique has been developed by adding thio compounds to alkali-ion based protocols and incubating at 45°C and the yield was more than 104 transformants/μg plasmid DNA.