Control of specific growth rate to enhance the production of a novel disintegrin, saxatilin, in recombinant Pichia pastoris.
The methylotrophic yeast Pichia pastoris is one of the best hosts for the production of foreign proteins because of the presence of a strong alcohol oxidase 1 (AOX1) promoter that can be induced by methanol. Feeding the yeast, methanol induces protein production and provides an energy source for the host cells. However, excessive levels of methanol inhibit the growth of host cells, and insufficient methanol levels lead to poor growth and protein production. We have used various methanol feeding strategies to enhance the production of saxatilin. Saxatilin is a novel snake venom-derived disintegrin that inhibits tumor angiogenesis and metastasis and has been shown to suppress ovarian cancer cell invasion. A two-step increase feeding strategy to control the specific growth rate led to the best results in terms of specific protein production rates and final saxatilin amounts within the limited fermentation time.
Min
Cheol-Ki
CK
Department of Integrated Biotechnology, Sogang University, Seoul 121-742, Republic of Korea.
Lee
Jin-Won
JW
Chung
Kwang-Hoe
KH
Park
Hong-Woo
HW
D019149:Bioreactors / Q000382:microbiology*; D049109:Cell Proliferation; D003470:Culture Media / Q000737:chemistry / Q000378:metabolism*; D019483:Disintegrins / Q000235:genetics / Q000378:metabolism*; D024861:Genetic Enhancement / Q000379:methods*; D010843:Pichia / Q000502:physiology*; D011995:Recombination, Genetic / Q000235:genetics
D003470:Culture Media; D019483:Disintegrins; C459080:saxatilin