Assessment of bacterial and fungal (hemi)cellulose-degrading enzymes in saccharification of ammonia fibre expansion-pretreated Arundo donax

Simona Giacobbe, Venkatesh Balan, Salvatore Montella, Massimo Fagnano, Mauro Mori, Vincenza Faraco

    Research output: Research - peer-reviewArticle

    • 1 Citations

    Abstract

    This study reports enzymatic hydrolysis of the biomass of the giant reed (Arundo donax L.) after ammonia fibre expansion (AFEX) pretreatment. In particular, the capacity of the arabinofuranosidase from the fungus Pleurotus ostreatus recombinantly expressed in Pichia pastoris rPoAbf, its evolved mutant rPoAbf F435Y/Y446F and the endo-cellulase from Streptomyces sp. G12 CelStrep recombinantly expressed in Escherichia coli to enhance the hydrolysis of AFEX-treated A. donax was investigated, using the corn stover as reference feedstock. The investigated enzymes were assayed using a mixture of purified cellulases (CBHI, CBHII, EGI and βG), endoxylanases (LX3, LX4) and accessory hemicellulases (LarbF and LβX) as reference enzyme mixture and substituting EGI with rCelStrep and LarbF with rPoAbf or rPoAbf F435Y/Y446F. The use of rPoAbf F435Y/Y446F in the substitution of LarbF led to improvements in sugar conversion, giving a glucan, xylan and arabinan conversion after 72 h of around 62, 63 and 80 %, respectively, similar or higher than those (44, 66 and 55 %) achieved by 72 h hydrolysis with commercial enzymes Novozymes Cellic®, Ctec3 and Htec3. The enzymes rPoAbf, rPoAbf F435Y/Y446F and rCelStrep were also investigated for their effect on hydrolysis of AFEX-pretreated A. donax by addition to commercial enzyme mixture Novozymes Cellic®, Ctec3 and Htec3, and it was shown that the addition of rPoAbf and its evolved mutant rPoAbf F435Y/Y446F enhanced both xylan and arabinan conversions, which achieved 80 % after 6 days of saccharification with rPoAbf F435Y/Y446F.

    LanguageEnglish (US)
    Pages2213-2224
    Number of pages12
    JournalApplied Microbiology and Biotechnology
    Volume100
    Issue number5
    DOIs
    StatePublished - Mar 1 2016

    Profile

    Poaceae
    Ammonia
    Cellulose
    Enzymes
    Hydrolysis
    Xylans
    araban
    Endo-1,4-beta Xylanases
    Pleurotus
    Cellulases
    Pichia
    Glucans
    Cellulase
    Streptomyces
    Biomass
    Zea mays
    Fungi
    Escherichia coli
    hemicellulase

    Keywords

    • Arabinofuranosidase
    • Cellulase
    • Lignocellulose
    • Pretreatment

    ASJC Scopus subject areas

    • Biotechnology
    • Applied Microbiology and Biotechnology

    Cite this

    Assessment of bacterial and fungal (hemi)cellulose-degrading enzymes in saccharification of ammonia fibre expansion-pretreated Arundo donax. / Giacobbe, Simona; Balan, Venkatesh; Montella, Salvatore; Fagnano, Massimo; Mori, Mauro; Faraco, Vincenza.

    In: Applied Microbiology and Biotechnology, Vol. 100, No. 5, 01.03.2016, p. 2213-2224.

    Research output: Research - peer-reviewArticle

    Giacobbe, Simona ; Balan, Venkatesh ; Montella, Salvatore ; Fagnano, Massimo ; Mori, Mauro ; Faraco, Vincenza. / Assessment of bacterial and fungal (hemi)cellulose-degrading enzymes in saccharification of ammonia fibre expansion-pretreated Arundo donax. In: Applied Microbiology and Biotechnology. 2016 ; Vol. 100, No. 5. pp. 2213-2224
    @article{b6a0f81b94e246aa8fe4d13e0a4d95f4,
    title = "Assessment of bacterial and fungal (hemi)cellulose-degrading enzymes in saccharification of ammonia fibre expansion-pretreated Arundo donax",
    abstract = "This study reports enzymatic hydrolysis of the biomass of the giant reed (Arundo donax L.) after ammonia fibre expansion (AFEX) pretreatment. In particular, the capacity of the arabinofuranosidase from the fungus Pleurotus ostreatus recombinantly expressed in Pichia pastoris rPoAbf, its evolved mutant rPoAbf F435Y/Y446F and the endo-cellulase from Streptomyces sp. G12 CelStrep recombinantly expressed in Escherichia coli to enhance the hydrolysis of AFEX-treated A. donax was investigated, using the corn stover as reference feedstock. The investigated enzymes were assayed using a mixture of purified cellulases (CBHI, CBHII, EGI and βG), endoxylanases (LX3, LX4) and accessory hemicellulases (LarbF and LβX) as reference enzyme mixture and substituting EGI with rCelStrep and LarbF with rPoAbf or rPoAbf F435Y/Y446F. The use of rPoAbf F435Y/Y446F in the substitution of LarbF led to improvements in sugar conversion, giving a glucan, xylan and arabinan conversion after 72 h of around 62, 63 and 80 %, respectively, similar or higher than those (44, 66 and 55 %) achieved by 72 h hydrolysis with commercial enzymes Novozymes Cellic®, Ctec3 and Htec3. The enzymes rPoAbf, rPoAbf F435Y/Y446F and rCelStrep were also investigated for their effect on hydrolysis of AFEX-pretreated A. donax by addition to commercial enzyme mixture Novozymes Cellic®, Ctec3 and Htec3, and it was shown that the addition of rPoAbf and its evolved mutant rPoAbf F435Y/Y446F enhanced both xylan and arabinan conversions, which achieved 80 % after 6 days of saccharification with rPoAbf F435Y/Y446F.",
    keywords = "Arabinofuranosidase, Cellulase, Lignocellulose, Pretreatment",
    author = "Simona Giacobbe and Venkatesh Balan and Salvatore Montella and Massimo Fagnano and Mauro Mori and Vincenza Faraco",
    year = "2016",
    month = "3",
    doi = "10.1007/s00253-015-7066-3",
    volume = "100",
    pages = "2213--2224",
    journal = "Applied Microbiology and Biotechnology",
    issn = "0175-7598",
    publisher = "Springer Verlag",
    number = "5",

    }

    TY - JOUR

    T1 - Assessment of bacterial and fungal (hemi)cellulose-degrading enzymes in saccharification of ammonia fibre expansion-pretreated Arundo donax

    AU - Giacobbe,Simona

    AU - Balan,Venkatesh

    AU - Montella,Salvatore

    AU - Fagnano,Massimo

    AU - Mori,Mauro

    AU - Faraco,Vincenza

    PY - 2016/3/1

    Y1 - 2016/3/1

    N2 - This study reports enzymatic hydrolysis of the biomass of the giant reed (Arundo donax L.) after ammonia fibre expansion (AFEX) pretreatment. In particular, the capacity of the arabinofuranosidase from the fungus Pleurotus ostreatus recombinantly expressed in Pichia pastoris rPoAbf, its evolved mutant rPoAbf F435Y/Y446F and the endo-cellulase from Streptomyces sp. G12 CelStrep recombinantly expressed in Escherichia coli to enhance the hydrolysis of AFEX-treated A. donax was investigated, using the corn stover as reference feedstock. The investigated enzymes were assayed using a mixture of purified cellulases (CBHI, CBHII, EGI and βG), endoxylanases (LX3, LX4) and accessory hemicellulases (LarbF and LβX) as reference enzyme mixture and substituting EGI with rCelStrep and LarbF with rPoAbf or rPoAbf F435Y/Y446F. The use of rPoAbf F435Y/Y446F in the substitution of LarbF led to improvements in sugar conversion, giving a glucan, xylan and arabinan conversion after 72 h of around 62, 63 and 80 %, respectively, similar or higher than those (44, 66 and 55 %) achieved by 72 h hydrolysis with commercial enzymes Novozymes Cellic®, Ctec3 and Htec3. The enzymes rPoAbf, rPoAbf F435Y/Y446F and rCelStrep were also investigated for their effect on hydrolysis of AFEX-pretreated A. donax by addition to commercial enzyme mixture Novozymes Cellic®, Ctec3 and Htec3, and it was shown that the addition of rPoAbf and its evolved mutant rPoAbf F435Y/Y446F enhanced both xylan and arabinan conversions, which achieved 80 % after 6 days of saccharification with rPoAbf F435Y/Y446F.

    AB - This study reports enzymatic hydrolysis of the biomass of the giant reed (Arundo donax L.) after ammonia fibre expansion (AFEX) pretreatment. In particular, the capacity of the arabinofuranosidase from the fungus Pleurotus ostreatus recombinantly expressed in Pichia pastoris rPoAbf, its evolved mutant rPoAbf F435Y/Y446F and the endo-cellulase from Streptomyces sp. G12 CelStrep recombinantly expressed in Escherichia coli to enhance the hydrolysis of AFEX-treated A. donax was investigated, using the corn stover as reference feedstock. The investigated enzymes were assayed using a mixture of purified cellulases (CBHI, CBHII, EGI and βG), endoxylanases (LX3, LX4) and accessory hemicellulases (LarbF and LβX) as reference enzyme mixture and substituting EGI with rCelStrep and LarbF with rPoAbf or rPoAbf F435Y/Y446F. The use of rPoAbf F435Y/Y446F in the substitution of LarbF led to improvements in sugar conversion, giving a glucan, xylan and arabinan conversion after 72 h of around 62, 63 and 80 %, respectively, similar or higher than those (44, 66 and 55 %) achieved by 72 h hydrolysis with commercial enzymes Novozymes Cellic®, Ctec3 and Htec3. The enzymes rPoAbf, rPoAbf F435Y/Y446F and rCelStrep were also investigated for their effect on hydrolysis of AFEX-pretreated A. donax by addition to commercial enzyme mixture Novozymes Cellic®, Ctec3 and Htec3, and it was shown that the addition of rPoAbf and its evolved mutant rPoAbf F435Y/Y446F enhanced both xylan and arabinan conversions, which achieved 80 % after 6 days of saccharification with rPoAbf F435Y/Y446F.

    KW - Arabinofuranosidase

    KW - Cellulase

    KW - Lignocellulose

    KW - Pretreatment

    UR - http://www.scopus.com/inward/record.url?scp=84958858727&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=84958858727&partnerID=8YFLogxK

    U2 - 10.1007/s00253-015-7066-3

    DO - 10.1007/s00253-015-7066-3

    M3 - Article

    VL - 100

    SP - 2213

    EP - 2224

    JO - Applied Microbiology and Biotechnology

    T2 - Applied Microbiology and Biotechnology

    JF - Applied Microbiology and Biotechnology

    SN - 0175-7598

    IS - 5

    ER -