bioprocessing in industrial organisms (such as Saccharomyces cerevisiae) as these hosts grow under conditions that are different from those where the enzymes
evolved (den Haan et al. 2015; la Grange et al. 2010).
Apart from gut fungal CAZymes, other proteins have been successfully cloned. Xylose
isomerase from Piromyces sp. E2 was successfully produced in Saccharomyces cerevisiae,
allowing it to grow on C5 as well as C6 sugars
(Kuyper et al. 2003). Swollenin from Pecoramyces ruminantium strain C1A was produced
in Escherichia coli and its addition to a lignocellulolytic enzyme cocktail enhanced overall
glucan and xylan hydrolysis yields from lignocellulosic biomass (Morrison et al. 2016b).
Finally, Haitjema (2017) produced fragments
of cellulosomal protein (ScaA) in Escherichia
coli and used ELISA assays to demonstrate its
strong binding activity to dockerin domains,
firmly suggesting that ScaA acts as a scaffolding
in these multi-enzyme complexes.
Table 17.2 Production of selected anaerobic fungal proteins in heterologous hosts
Activity
Species/strain
Expression host
References
Endoglucanase (endo1,4-b-glucanase)
Neocallimastix
patriciarum
Clostridium beijerinckii
NCIMB 8052
Lo ´pez-Contreras et al.
(2001)
Cellobiohydrolase
Orpinomyces sp.Y102
Escherichia coli
Chen et al. (2014)
b-D-glucosidase
Orpinomyces sp. PC-2
Saccharomyces cerevisiae
Li et al. (2004)
Piromyces sp. strain E2 Pichia pastoris
Harhangi et al. (2002)
Saccharomyces cerevisiae
O’Malley et al. (2012)
Xylanases (endo-b 1 ,4xylanase)
Neocallimastix frontalis Kluyveromyces lactis and
Penicillium roqueforti
Durand et al. (1999)
Orpinomyces sp. PC-2
Hypocrea jecorina
Li et al. (2007)
B-xylosidase
Pecoramyces
ruminantium
strain C1A
Escherichia coli
Morrison et al. (2016a)
Xylose isomerase
Piromyces sp. strain E2 Saccharomyces cerevisiae
Kuyper et al. (2003);
O’Malley et al. (2012)
Xylulokinase
Piromyces sp. strain E2 Saccharomyces cerevisiae
O’Malley et al. (2012)
Glycosyl hydrolase 48
Piromyces sp. strain E2 Saccharomyces cerevisiae
O’Malley et al. (2012)
Fungal Scaffoldin (ScaA) Piromyces finnis
Anaeromyces
robustus
Neocallimastix
californiae
Escherichia coli
Haitjema et al. (2017)
Acetyl xylan esterase
Neocallimastix
patriciarum
Escherichia coli
Dalrymple et al. (1997)
Orpinomyces sp. PC-2
Escherichia coli
Blum et al. (1999)
Feruloyl esterase
Anaeromyces
mucronatus
Escherichia coli
Gruninger et al. (2016)
Piromyces equi
Trichoderma reesei
Poidevin et al. (2009)
Mannanase
Piromyces sp.
Escherichia coli
Ali et al. (1995)
Galacto-(a-1,2)arabinosidase
Neocallimastix frontalis Escherichia coli
Jones et al. (2017)
b-1,2-arabinobiosidase
Neocallimastix frontalis Escherichia coli
Jones et al. (2017)
Lichenase
Orpinomyces sp. PC-2
Escherichia coli
Chen et al. (1997)
Swollenin
Pecoramyces
ruminantium
strain C1A
Escherichia coli
(Morrison et al. 2016b)
Celpin
Piromyces sp. strain E2 Saccharomyces cerevisiae
O’Malley et al. (2012)
Cylophillin
Orpinomyces sp. PC-2
Escherichia coli
Chen et al. (1995); Chen
et al. (2006)
CBM29
Piromyces equi
a
Nicotiana tabacum
Obembe et al. (2007)
a
Proper taxonomic description is missing for this isolate
422
V. Flad et al.
evolved (den Haan et al. 2015; la Grange et al. 2010).
Apart from gut fungal CAZymes, other proteins have been successfully cloned. Xylose
isomerase from Piromyces sp. E2 was successfully produced in Saccharomyces cerevisiae,
allowing it to grow on C5 as well as C6 sugars
(Kuyper et al. 2003). Swollenin from Pecoramyces ruminantium strain C1A was produced
in Escherichia coli and its addition to a lignocellulolytic enzyme cocktail enhanced overall
glucan and xylan hydrolysis yields from lignocellulosic biomass (Morrison et al. 2016b).
Finally, Haitjema (2017) produced fragments
of cellulosomal protein (ScaA) in Escherichia
coli and used ELISA assays to demonstrate its
strong binding activity to dockerin domains,
firmly suggesting that ScaA acts as a scaffolding
in these multi-enzyme complexes.
Table 17.2 Production of selected anaerobic fungal proteins in heterologous hosts
Activity
Species/strain
Expression host
References
Endoglucanase (endo1,4-b-glucanase)
Neocallimastix
patriciarum
Clostridium beijerinckii
NCIMB 8052
Lo ´pez-Contreras et al.
(2001)
Cellobiohydrolase
Orpinomyces sp.Y102
Escherichia coli
Chen et al. (2014)
b-D-glucosidase
Orpinomyces sp. PC-2
Saccharomyces cerevisiae
Li et al. (2004)
Piromyces sp. strain E2 Pichia pastoris
Harhangi et al. (2002)
Saccharomyces cerevisiae
O’Malley et al. (2012)
Xylanases (endo-b 1 ,4xylanase)
Neocallimastix frontalis Kluyveromyces lactis and
Penicillium roqueforti
Durand et al. (1999)
Orpinomyces sp. PC-2
Hypocrea jecorina
Li et al. (2007)
B-xylosidase
Pecoramyces
ruminantium
strain C1A
Escherichia coli
Morrison et al. (2016a)
Xylose isomerase
Piromyces sp. strain E2 Saccharomyces cerevisiae
Kuyper et al. (2003);
O’Malley et al. (2012)
Xylulokinase
Piromyces sp. strain E2 Saccharomyces cerevisiae
O’Malley et al. (2012)
Glycosyl hydrolase 48
Piromyces sp. strain E2 Saccharomyces cerevisiae
O’Malley et al. (2012)
Fungal Scaffoldin (ScaA) Piromyces finnis
Anaeromyces
robustus
Neocallimastix
californiae
Escherichia coli
Haitjema et al. (2017)
Acetyl xylan esterase
Neocallimastix
patriciarum
Escherichia coli
Dalrymple et al. (1997)
Orpinomyces sp. PC-2
Escherichia coli
Blum et al. (1999)
Feruloyl esterase
Anaeromyces
mucronatus
Escherichia coli
Gruninger et al. (2016)
Piromyces equi
Trichoderma reesei
Poidevin et al. (2009)
Mannanase
Piromyces sp.
Escherichia coli
Ali et al. (1995)
Galacto-(a-1,2)arabinosidase
Neocallimastix frontalis Escherichia coli
Jones et al. (2017)
b-1,2-arabinobiosidase
Neocallimastix frontalis Escherichia coli
Jones et al. (2017)
Lichenase
Orpinomyces sp. PC-2
Escherichia coli
Chen et al. (1997)
Swollenin
Pecoramyces
ruminantium
strain C1A
Escherichia coli
(Morrison et al. 2016b)
Celpin
Piromyces sp. strain E2 Saccharomyces cerevisiae
O’Malley et al. (2012)
Cylophillin
Orpinomyces sp. PC-2
Escherichia coli
Chen et al. (1995); Chen
et al. (2006)
CBM29
Piromyces equi
a
Nicotiana tabacum
Obembe et al. (2007)
a
Proper taxonomic description is missing for this isolate
422
V. Flad et al.