Method for synthesizing 11alpha, 15alpha-diOH-4-AD through Gibberella intermedia CA3-1
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A technology of -dioh-4-ad and 4-AD, applied in the field of androstenedione as 11α, 15α-diOH-4-AD
Inactive Publication Date: 2016-03-23
JIANGNAN UNIV
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[0015] (1) Prepare the seed liquid of Gibberella intermedia CA3-1 bacterial strain
[0016] Pick a loop of Gibberella intermedia CA3-1 strain on the solid PDA medium, inoculate it in a 500mL Erlenmeyer flask containing 100mL seed medium, place it on a shaker at 200r / min at 30°C Cultivate for 20-24 hours to the logarithmic phase, and the seed solution of CA3-1 is obtained;
[0017] (2) Insert the seed culture solution into a 250mL shake flask equipped with 30mL fermentation medium with an inoculum size of 4%, and continue to cultivate for 20 hours under the same conditions to obtain a bacterial cell fermentation solution;
[0018] (3) Put 4-AD into the bacterial fermentation broth, the feeding concentration is 4.0g / L, transform under the same conditions for 60 hours, extract the product for HPLC analysis, the calculated conversion rate of 4-AD is 99.3%, 11α, 15α The yield of -diOH-4-AD w...
Embodiment 2
[0020] (1) Seed culture and fermentation culture are the same as in Example 1.
[0021] (2) Put 4-AD into the bacterial fermentation broth, the feeding concentration is 6.0g / L, transform under the same conditions for 60 hours, extract the product for HPLC analysis, the calculated conversion rate of 4-AD is 80.4%, 11α, 15α The yield of -diOH-4-AD was 52.3%.
Embodiment 3
[0023] (1) Seed culture and fermentation culture are the same as in Example 1.
[0024] (2) Put 4-AD into the bacterial fermentation broth, the feeding concentration is 8.0g / L, transform under the same conditions for 60 hours, extract the product for HPLC analysis, the calculated conversion rate of 4-AD is 61.3%, 11α, 15α The yield of -diOH-4-AD was 41.7%.
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Abstract
The invention belongs to the biotechnology field, and particularly relates to a method for converting a substrate 4-AD into 11alpha, 15alpha-diOH-4-AD through Gibberella intermedia CA3-1. The bacterial strain is preserved in the Common Microorganism Center of China Committee for Culture Collection of Microorganisms, the preservation number is CGMCC No. 4903. The 4-AD can be converted into 11alpha, 15alpha-diOH-4-AD through the Gibberella intermedia CA3-1; when the substrate feed concentration is 4 g/L, the 4-AD conversion rate reaches up to 99.3%, and the yield of the 11alpha, 15alpha-diOH-4-AD is 68.7%. The synthesis of the 11alpha, 15alpha-diOH-4-AD provides a more diversified precursor for research of steroid drugs and intermediates thereof, and the certain research value and market significance are achieved.
Description
technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for converting a substrate androstenedione (4-AD) into 11α, 15α-diOH-4-AD by using a strain of Gibberella intermedia CA3-1. Background technique [0002] Androstenedione (4-AD) is an important intermediate of steroid hormone drugs, which can be used to synthesize intermediates of various steroid hormone drugs such as testosterone, methyltestosterone, estrone and adrenocortical hormone . It can be oxidized in vivo to produce various hydroxylated products such as 9α-OH-4-AD, 11α-OH-4-AD and 11α,15α-diOH-4-AD, among which 9α-OH-4-AD There are many reports. 9α-OH-4-AD can be used to synthesize corticosteroids, antiandrogen, antiestrogens and contraceptive drugs, and is a very important steroid hormone intermediate. However, there are few reports about its 11α and 15α dihydroxy products. Compared with traditional chemical synthesis methods, microbial transformatio...
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