title: Effect of codon optimisation on the production of recombinant fish growth hormone in Pichia pastoris.
abstract: This study was established to test the hypothesis of whether the codon optimization of fish growth hormone gene (FGH) based on P. pastoris preferred codon will improve the quantity of secreted rFGH in culture supernatant that can directly be used as fish feed supplements. The optimized FGH coding sequence (oFGH) and native sequence (nFGH) of giant grouper fish (Epinephelus lanceolatus) were cloned into P. pastoris expression vector (pPICZαA) downstream of alcohol oxidase gene (AOX1) for efficient induction of extracellular rFGH by adding 1% of absolute methanol. The results showed that recombinant P. pastoris was able to produce 2.80 ± 0.27 mg of oFGH compared to 1.75 ± 0.25 of nFGH in one litre of culture supernatant. The total body weight of tiger grouper fingerlings fed with oFGH increased significantly at third (P < 0.05) and fourth weeks (P < 0.01) of four-week experiment period compared to those fed with nFGH. Both oFGH and nFGH significantly enhanced the final biomass and fish survival percentage. In conclusion, codon optimization of FGH fragment was useful to increase rFGH quantity in the culture supernatant of P. pastoris that can be directly used as fish feed supplements. Further studies are still required for large scale production of rFGH and practical application in aquaculture production.
authors.affiliation: Institute of Research Management & Monitoring, Deputy Vice Chancellor (Research & Innovation) Building, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: Hussin A
authors.initials: HA
authors.lastname: Rothan
authors.affiliation: Genetics and Molecular Biology Unit, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: Teh Ser
authors.initials: TS
authors.lastname: Huy
authors.affiliation: Genetics and Molecular Biology Unit, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: Zulqarnain
authors.initials: Z
authors.lastname: Mohamed
mesh_terms: D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D005399:Fishes / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D013006:Growth Hormone / Q000096:biosynthesis* / Q000235:genetics*; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins; D017422:Sequence Analysis, DNA
chemical_list: D003062:Codon; D011994:Recombinant Proteins; D013006:Growth Hormone