From mannan to bioethanol: cell surface co-display of β-mannanase and β-mannosidase on yeast Saccharomyces cerevisiae.
BACKGROUND
Mannans represent the largest hemicellulosic fraction in softwoods and also serve as carbohydrate stores in various plants. However, the utilization of mannans as sustainable resources has been less advanced in sustainable biofuel development. Based on a yeast cell surface-display technology that enables the immobilization of multiple enzymes on the yeast cell walls, we constructed a recombinant Saccharomyces cerevisiae strain that co-displays β-mannanase and β-mannosidase; this strain is expected to facilitate ethanol fermentation using mannan as a biomass source.


RESULTS
Parental yeast S. cerevisiae assimilated mannose and glucose as monomeric sugars, producing ethanol from mannose. We constructed yeast strains that express tethered β-mannanase and β-mannosidase; co-display of the two enzymes on the cell surface was confirmed by immunofluorescence staining and enzyme activity assays. The constructed yeast cells successfully hydrolyzed 1,4-β-d-mannan and produced ethanol by assimilating the resulting mannose without external addition of enzymes. Furthermore, the constructed strain produced ethanol from 1,4-β-d-mannan continually during the third batch of repeated fermentation. Additionally, the constructed strain produced ethanol from ivory nut mannan; ethanol yield was improved by NaOH pretreatment of the substrate.


CONCLUSIONS
We successfully displayed β-mannanase and β-mannosidase on the yeast cell surface. Our results clearly demonstrate the utility of the strain co-displaying β-mannanase and β-mannosidase for ethanol fermentation from mannan biomass. Thus, co-tethering β-mannanase and β-mannosidase on the yeast cell surface provides a powerful platform technology for yeast fermentation toward the production of bioethanol and other biochemicals from lignocellulosic materials containing mannan components.
Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501 Japan.
Jun
J
Ishii
Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501 Japan.|Department of Life Sciences, Graduate School of Bioresources, Mie University, 1577, Kurimamachiya, Tsu, Mie 514‑8507 Japan.
Fumiyoshi
F
Okazaki
Research Center for Biotechnology, Indonesian Institute of Sciences (LIPI), Cibinong Jalan Raya Bogor Km. 46, Cibinong, West Java 16911 Indonesia.
Apridah Cameliawati
AC
Djohan
Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501 Japan.|Department of Environmental Sciences, Graduate School of Nutritional and Environmental Sciences, University of Shizuoka, 52-1 Yada, Suruga, Shizuoka 422-8526 Japan.
Kiyotaka Y
KY
Hara
Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501 Japan.
Nanami
N
Asai-Nakashima
Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501 Japan.
Hiroshi
H
Teramura
Research Center for Biotechnology, Indonesian Institute of Sciences (LIPI), Cibinong Jalan Raya Bogor Km. 46, Cibinong, West Java 16911 Indonesia.
Ade
A
Andriani
Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501 Japan.
Masahiro
M
Tominaga
Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501 Japan.
Satoshi
S
Wakai
Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501 Japan.
Prihardi
P
Kahar
Research Center for Biotechnology, Indonesian Institute of Sciences (LIPI), Cibinong Jalan Raya Bogor Km. 46, Cibinong, West Java 16911 Indonesia.
Yopi
Research Center for Biotechnology, Indonesian Institute of Sciences (LIPI), Cibinong Jalan Raya Bogor Km. 46, Cibinong, West Java 16911 Indonesia.
Bambang
B
Prasetya
Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501 Japan.
Chiaki
C
Ogino
Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501 Japan.|Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501 Japan.|RIKEN Center for Sustainable Resource Science, 1-7-22, Suehiro, Tsurumi, Yokohama, 230-0045 Japan.
Akihiko
A
Kondo
Biofuel; Biomass resource; Cell surface display; Ethanol; Fermentation; Mannan; Mannanase; Mannosidase; Saccharomyces cerevisiae; Yeast