A novel host-vector system for heterologous protein co-expression and purification in the Trichoderma reesei industrial strain RUT-C30.
OBJECTIVE
To develop a host-vector system for heterologous protein co-expression and purification in the Trichoderma reesei (teleomorph Hypocrea jecorina) industrial strain RUT-C30.


RESULTS
The co-expression and purification system is based on: (i) an efficient and reliable selectable marker using bar (phosphinothricin acetyltransferase gene); (ii) a compact hygromycin B resistance marker; and (iii) a versatile integration plasmid for gene expression and knockout. Fluorescent protein genes were introduced into T. reesei and the corresponding proteins were purified from fermentation broth. The host-vector system was used in a proof-of-principle approach to achieve the co-expression of an alkaline endoglucanase and an alkaline cellobiohydrolase.


CONCLUSIONS
This protocol can be used at an industrial scale to produce large amounts of proteins in T. reesei.
State Key Lab of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Meilong Road, P.O.B. 311, 130, Shanghai, 200237, China.
Lei
L
Zhang
Research Center for Biomass Energy, East China University of Science and Technology, Shanghai, 200237, China.
Suping
S
Zhang
Tianjin Hualida Biotechnology Co. Ltd., No. 203, Huanghai Rd, TEDA, Tianjin, 300457, China.
Xin
X
Jiang
State Key Lab of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Meilong Road, P.O.B. 311, 130, Shanghai, 200237, China.
Wei
W
Wei
State Key Lab of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Meilong Road, P.O.B. 311, 130, Shanghai, 200237, China. wadexp@ecust.edu.cn.
Wei
W
Wang
State Key Lab of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Meilong Road, P.O.B. 311, 130, Shanghai, 200237, China. dzhwei@ecust.edu.cn.
Dongzhi
D
Wei
D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism; D025141:Drug Resistance, Fungal; D005285:Fermentation; D006921:Hygromycin B / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014242:Trichoderma / Q000235:genetics* / Q000378:metabolism
D011994:Recombinant Proteins; D006921:Hygromycin B; D000123:Acetyltransferases; C056243:phosphinothricin N-acetyltransferase
Alkaline cellulose; Co-expression; Filamentous fungi; Phosphinothricin; Trichoderma reesei; xyn2 Promoter