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Volume 7, Issue 3(Suppl) J Biotechnol Biomater, an open access journal
ISSN: 2155-952X Euro Biotechnology 2017
September 25-27, 2017
17th EURO BIOTECHNOLOGY CONGRESS
September 25-27, 2017 Berlin, Germany
Hyper cellulase-producing fungus Talaromyces pinophilus EMMdevelopment through randommutagenesis and
genetic engineering
Anli Geng1, Zunsheng Wang1, Rupali Rahul Manglekar1, Fen Liu1, 2, Zhiyi Zhou1, 2, Huirong Zhang1, 2 and Youhong Zhang2
1Ngee Ann Polytechnic, Singapore
2Wuhan Institute of Technology, China
Talaromyces pinophilus UTA1 and EMM are cellulase hyper-producing mutants that originated from T. pinophilus OPC4-1 through
UV irradiation and chemical mutagenesis by NTG and EMS. Full genome sequencing of these two mutants and the parent
strain was conducted and 73 genes were identified with either SNPs or InDels. Functions of the 73 genes were identified using NCBI
GenBank database. Among the 73 genes, 3 transcription factors were identified. They might be responsible for the enhancement of
cellulase activity in mutant strains, UTA1 and EMM. Genes encoding the 3 transcription factors were successfully cloned to further
confirm their enhancement in cellulase and hemicellulase production in mutant strains. Further genetic engineering of the mutant
strain EMM was conducted to further enhance its enzyme production. A uracil auxotroph strain T. pinophilus EMU was isolated
through random mutagenesis. A wild-type pyrF gene encoding orotate phosphoribosyl transferase (OPRTase, EC 2.4.2.10) isolated
from T. pinophilus OPC4-1, the parent strain can be used as the selection marker for genetic engineering of strain T. pinophilus EMM.
A marker recycle system was developed and was used for the knock-out of creA gene, the gene mediating catabolite repression. A
creA gene knock-out strain, A creA 21 was successfully isolated. It demonstrated enhanced cellulase and xylanase production and
higher resistance to the increased glucose concentration. The genetic engineering tools were successfully developed for strain T.
pinophilus EMM and disruption of creA gene in strain EMM was effective for enhanced enzyme production.
Biography
Anli Geng is currently an Assistant Director of Life Sciences and Chemical Technology of Ngee Ann Polytechnic. She currently holds the President position in
BioEnergy Society of Singapore (BESS). She is also the Co-founder and Director of Sunvisiae Biotech Pte Ltd, a Singapore-based industrial biotechnology
company. Prior to joining Ngee Ann Polytechnic, she was working at Institute of Environmental Science and Engineering (IESE) as a Research Scientist. She
has more than 25 years of R&D experience, working extensively in environmental biotechnology, green energy technology and industrial biotechnology. She
has more than 30 journal publications and her work has been presented in many international conferences. Her current research focuses on developing novel
microorganisms to produce industrial enzymes, chemicals and fuels, novel nutraceuticals and cosmetics ingredients at Ngee Ann Polytechnic. She obtained Ngee
Ann Polytechnic Staff Excellence Award and IChemE Award on Sustainable Technology in 2012.
Anli Geng et al., J Biotechnol Biomater 2017, 7:3(Suppl)
DOI: 10.4172/2155-952X-C1-076
Figure 1: Zone of clearance generated by creA knock-out mutant of T. pinophilus EMM