title: Enhanced porcine circovirus Cap protein production by Pichia pastoris with a fuzzy logic DO control based methanol/sorbitol co-feeding induction strategy.
abstract: Porcine circovirus Cap protein production by P. pastoris with strong AOX promoter suffered with the problems with traditional pure methanol induction: (1) inefficient methanol metabolism; (2) extensive oxygen supply load; (3) difficulty in stable DO control; (4) low protein titer. In this study, based on the difference of DO change patterns in response to methanol and sorbitol additions, a novel fuzzy control system was proposed to automatically regulate the co-feeding rates of methanol and sorbitol for efficient Cap protein induction. With aid of the proposed control system when setting DO control level at 10%, overall fermentation performance was significantly improved: (1) DO could be stably controlled under mild aeration condition; (2) methanol consumption rate could be restricted at moderate level and the major enzymes involved with methanol metabolism were largely activated; (3) Cap protein concentration reached a highest level of 198mg/L, which was about 64% increase over the best one using the pure methanol induction strategies.
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Ding
authors.affiliation: Institute of Animal Husbandry and Veterinary Research, Shanghai Academy of Agricultural Science, Shanghai 201106, China. Electronic address: clzhang201310@163.com.
authors.forename: Chunling
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
authors.forename: Minjie
authors.initials: M
authors.lastname: Gao
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
authors.forename: Guoli
authors.initials: G
authors.lastname: Hou
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
authors.forename: Kexue
authors.initials: K
authors.lastname: Liang
authors.affiliation: Institute of Animal Husbandry and Veterinary Research, Shanghai Academy of Agricultural Science, Shanghai 201106, China.
authors.forename: Chunhua
authors.initials: C
authors.lastname: Li
authors.affiliation: Institute of Animal Husbandry and Veterinary Research, Shanghai Academy of Agricultural Science, Shanghai 201106, China.
authors.forename: Jianping
authors.initials: J
authors.lastname: Ni
authors.affiliation: Institute of Animal Husbandry and Veterinary Research, Shanghai Academy of Agricultural Science, Shanghai 201106, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Li
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China. Electronic address: zpshi@jiangnan.edu.cn.
authors.forename: Zhongping
authors.initials: Z
authors.lastname: Shi
mesh_terms: D000818:Animals; D019149:Bioreactors / Q000382:microbiology; D017925:Circovirus / Q000201:enzymology*; D005285:Fermentation; D017143:Fuzzy Logic; D000432:Methanol / Q000378:metabolism*; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics; D013012:Sorbitol / Q000378:metabolism*; D013552:Swine; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins; D014764:Viral Proteins; D013012:Sorbitol; D000432:Methanol
keywords: Cap protein; DO; Fermentation; Fuzzy control; Metabolic analysis