Effects of soybean oil and alanine on rifamycin SV biosynthesis in 250 mL flask were studied in this paper. When 3 mL/L soybean oil and 0.8 g/L alanine was added into the medium at 72 h, rifamycin SV production was increased by 5.3% and 6.9% over the control, respectively. Response surface methodology was used to study the interaction between soybean oil and alanine. Result showed that their optimal amount was 2.57 mL/L and 0. 89 g/ L, respectively. Under the optimal conditions, rifamycin SV production reached 5179 U/mL that was 12.3% higher than control. The results of organic acids analysis demonstrated that utilization rate of a-ketoglutarate and citriate was 0.86 mg/（L · h） and 0.77 mg/（L · h） , respectively, which was 115.7% and 23.3% higher than that of control in the fermentation process from 72 h to120 h. Meanwhile, succinate concentration reached 1.01 g/L that was 16.1% higher than the control. The result indicated that the conversion efficiency of α-ketoglutarate and citrate to succinate in the TCA cycle was greatly improved with the addition of oil and alanine, which led to enhanced synthesis of succinyl- CoA and methylmalony-CoA and the facilitation of rifamycin SV biosynthesis.%研究了在250mL摇瓶发酵过程中添加豆油和丙氨酸对利福霉素SV生物合成的影响。72h分别向培养基中添加3mL／L豆油和0．8g／L丙氨酸，发酵液中利福霉素SV的效价分别比对照提高5．3％和6．9％。通过响应面分析法优化考察了豆油和丙氨酸对利福霉素SV合成的交互作用，确定在72h添加0．89g／L的丙氨酸和2．57mL／L豆油时，摇瓶发酵单位是5179U／mL，比对照提高12．3％。有机酸分析结果显示，发酵培养72—120h，添加组的α-酮戊二酸和柠檬酸的利用速率分别是0．86mg／（L·h）和0．77mg
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