Improvement of penicillin yields in solid-state and submerged fermentation of Penicillium chrysogenum by amplification of the penicillin biosynthetic gene cluster |
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Authors: | Carolina Campos Francisco J Fernández Edgar C Sierra Francisco Fierro Adriana Garay Javier Barrios-González |
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Institution: | 1. Departamento de Ciencias de la Salud, Universidad Autónoma Metropolitana, Iztapalapa, Mexico D.F., Mexico 2. Departamento de Biotecnología, Universidad Autónoma Metropolitana—Iztapalapa, Av. San Rafael Atlixco 186, Col. Vicentina, Iztapalapa, 09340, Mexico D.F., Mexico 3. Academia de Biología, Universidad Autónoma de la Ciudad de México, Mexico, Mexico 4. Departamento de Producción Agrícola y Animal, Universidad Autónoma Metropolitana, Xochimilco, Mexico D.F., Mexico 5. Genética Molecular, Desarrollo y Evolución de Plantas, Inst. de Ecología, Universidad Nacional Autónoma de México, 3er Circuito Ext. s/no. C.U., 04510, México D.F., Mexico
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Abstract: | Penicillium chrysogenum low and high penicillin producing strains were transformed with a cosmid containing the whole penicillin biosynthetic gene
cluster. The cosmid library was constructed in a newly developed cosmid vector, IztapaCos, which allows cloning and direct
introduction of large DNA fragments in fungal recipients using phleomycin resistance as selection marker. The effect of increased
gene dosage on penicillin production was evaluated both in submerged (SmF) and solid-state fermentation (SSF). Transformants
from the low-producing strain Wis 54-1255, showed a 67.3 and 28.3% increased penicillin titer in SSF and SmF, respectively.
In transformants from the high-producing strain P2-32 the increase was 92.9 and 158.4% respectively. Strain P2-32 already
contains originally about 14 copies of the penicillin biosynthetic cluster, which shows that the strategy of increasing the
gene dosage is still valid for high copy-number strains. The different behavior of the two strains in each type of culture
is discussed, along with the practical implications for industrial penicillin production. |
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Keywords: | Penicillin gene cluster amplification Penicillin production increase Submerged and solid-state fermentation |
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