生物工程學(xué)報 Chin J Biotech 2008, January 25; 24(1): 15-20

摘 要: 在格爾德霉素產(chǎn)生菌吸水鏈霉菌17997(Streptomyces hygroscopicus 17997)中存在兩種3-氨基-5-羥基苯甲酸(3-amino-5-hydroxybenzoic acid, AHBA)的生物合成基因簇, 根據(jù)同源性可分為苯醌類和萘醌類,。已證明其中苯醌類的AHBA生物合成基因簇負(fù)責(zé)格爾德霉素(geldanamycin, Gdm)起始單位的合成, 而萘醌類的AHBA基因簇可能參與未知安莎化合物的生物合成。為提高吸水鏈霉菌17997菌種的Gdm發(fā)酵產(chǎn)量, 并研究高產(chǎn)菌種在固體培養(yǎng)基上孢子的生長周期,。采用基因阻斷技術(shù), 將吸水鏈霉菌17997中的萘醌類AHBA生物合成基因簇(shnSOP)進(jìn)行破壞, 以獲得ΔSOP菌株, 從而減少對合成所需共同底物AHBA的爭奪,。HPLC分析結(jié)果表明ΔSOP菌株Gdm的發(fā)酵產(chǎn)量比原株提高185%。同時, 通過孢子計數(shù)發(fā)現(xiàn)該菌株在固體培養(yǎng)基上的孢子生長經(jīng)歷2個周期, 第2代孢子菌種的Gdm產(chǎn)量較高,。
關(guān)鍵詞: 吸水鏈霉菌17997, 格爾德霉素, 基因阻斷, 孢子形成周期

Construction and Cultivation of Genetically-engineered Strain to Improve Geldanamycin Production
Weiqing He, Hongxia Zhou, Hongyuan Wang, Qunjie Gao, and Yiguang Wang
Institute of Medicinal Biotechnology, CAMS & PUMC, Key Laboratory of Biotechnology of Antibiotics, Ministry of Health, Beijing 100050, China

Abstract: To improve the production of geldanamycin in Streptomyces hygroscopicus 17997, gene disruption was done to delete the naphthalenic AHBA genes (shnSOP), encoding the products that share the common biosynthetic substrates with geldanamycin. The resulting mutant strain (ΔSOP) was cultivated on a solid medium and the amount of spores collected from the plates was calculated from 5 to 14 days and the yield of geldanamycin was measured by HPLC. The geldanamycin production of the ΔSOP strain increased by 185% comparing with that of the parent strain. On solid medium, the ΔSOP strain underwent 2 cycles of sporulation and the growth of the second sporulation had the highest geldanamycin production.
Keywords: Streptomyces hygroscopicus17997, geldanamycin, gene disruption, spore formation

全文鏈接：格爾德霉素基因工程高產(chǎn)菌株的構(gòu)建和培養(yǎng)

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