Aspergillus niger strain and its culture method and application

A cultivation method, the technology of Aspergillus niger, which is applied in the direction of fungi, hydrolytic enzymes, fermentation, etc., can solve many problems, and achieve the effect of fast growth, not easy to mutate, and simple cultivation method

Inactive Publication Date: 2004-06-30
SHANGHAI INST OF ORGANIC CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] There are few reports on the asymmetric hydrolys

Method used

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  • Aspergillus niger strain and its culture method and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] The cultivation of embodiment 1 thalline (1)

[0017] Use an inoculation loop to pick Aspergillus niger CGMCC 0496 thallus and connect it to the above-mentioned slant, grow in medium at 10-30°C for 2-3 days, and a large number of black spore groups appear. Then pick the well-grown bacteria from the slope and put them into the culture medium, culture them on a reciprocating shaker at 10-30°C for 2-3 days, centrifuge the obtained bacteria (5,000rmp) for 30 minutes, and wash the bacteria with 0.8% normal saline Wash, then centrifuge or filter to obtain the bacterial cells, and the obtained bacterial cells can be directly used for the reaction. The bacteria are gray spherical.

[0018] The slant medium contains 10-30g / L fructose, glucose or sucrose, 6-15g / L corn steep liquor, yeast extract or corn flour, 1-3g / L agar, 1.0g / L K 2 HPO 4 ·H 2 O, 0.5g / L KCl, 0.5g / L MgSO 4 ·H 2 O, 0.01g / L FeSO 4 ·H 2 O, adjust the pH value to 7 with inorganic bases such as ...

Embodiment 2

[0021] Example 2 Asymmetric Resolution of p-Chlorostyrene Oxirane

[0022] The product (S)-3 was obtained with a yield of 40% using p-chlorostyrene oxide as a substrate. Product (R)-4, yield 44%.

[0023] Product (S)-3: Optical purity: 96% [α] D 25 +19.2°(c1.0, CHCl 3 )

[0024] 1 NMR (CCl 4 +TMS, 60MHz) δ=2.6(dd, J 1 =6Hz,J 2 =2Hz, 1H), δ=3.1(dd, J 1 =6Hz,J 2 =4Hz, 1H), δ=3.6-3.8(m, 1H), δ=6.9-7.4(m, 4H)

[0025] MS m / z(rel.intensity%)156(M + +2, 2), 155 (M + +1, 3), 154 (M + , 8), 138(3), 125(40), 119(39), 91(29), 89(100)

[0026] Product (R)-4: Optical purity: 87% [G] D 25 -33.2° (c1.0, EtOH)

[0027] 1 NMR (CD 3 COCD 3+TMS, 300MHz) δ=3.13(bs, 2H), δ=3.53(dd, J 1 =10.9Hz,J 2 =7.4Hz, 1H), δ=3.61(dd, J 1 =11Hz,J 2 =4.6, 1H), δ=4.71(dd, J 1 =7.3Hz,J 2 =4.5Hz, 1H), δ=7.12-7.52(m, 4H)

[0028] MS m / z(rel.instensity%)172(M + , 2), 155(11), 141(86), 125(8), 113(26), 77(100), 51(21)

Embodiment 3

[0029] Example 3 Asymmetric resolution of p-bromostyrene oxide

[0030] The product (S)-5 was obtained with p-bromostyrene oxide as the substrate, and the yield was 35%. Product (R)-6, yield 43%.

[0031] Product (S)-5: Optical purity: 100% [α] D 25 +14.2° (c1.5, CHCl 3 )

[0032] 1 NMR (CCl 4 +TMS, 90MHz) δ=2.6(dd, J 1 =5Hz,J 2 =2Hz, 1H), δ=3.0(dd, J 1 =J 2 =4.5Hz, 1H), δ=3.6-3.7(m, 1H), δ=7.1-7.4(AB, J=6Hz, 4H)

[0033] MS m / z(rel.instensity%)200(M + +2, 3), 199 (M + +1, 3), 198 (M + , 4), 197(M+-1, 3), 169(14), 119(41), 89(100), 63(36)

[0034] Product (R)-6: Optical purity: 87% [α] D 25 -38.4° (c1.0, CHCl 3 )

[0035] 1 NMR (CD 3 COCD 3 +TMS, 300MHz) δ=3.58(bs, 2H), δ=3.88-3.94(m, 1H), δ=3.48(m, 1H), δ=4.71(m, 1H), δ=7.36-7.48(AB , J=8.4Hz, 4H)

[0036] MS m / z (rel.intensity %) 218, 216 (M + , 3), 187(71), 185(83), 159(16), 157(19), 77(100), 51(18)

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Abstract

An Asperillus niger 5450 strain (CGMCCNO.0496) is disclosed, which is obtained by screening from soil and well grows in culture medium and culture liquid. After growing in culture liquid for 2-3 days, it has very high reaction activity and contains epoxy hydrolase with corresponding selectivity. It can be directly used in asymmetric hydrolysis of recemic 1,2-epoxide to obtain S-type epoxide and R-type 1.2-diol.

Description

technical field [0001] The invention relates to a microbial catalyst and its cultivation method Background technique [0002] Optically active 1,2-epoxides and their corresponding diols have extremely high application value in pesticides, medicine and fine chemicals. Such as: Calcium ion channel blockers (a class of cardiovascular drugs), according to Press release, Zeneca, 1996 speculation: the world market share reaches 4 billion US dollars, the US market accounts for 2 billion, of which β-receptor blockers The annual sales volume of pharmaceuticals in the United States exceeds 350 million US dollars, accounting for 22% of the antitanginal market in the United States [Pharmaprojects CD v2.1, copyright 1999 PJB Publication Ltd.Richmond, Surrey, UK.]. Optically active epoxy and diols are the key intermediates for the synthesis of cardiovascular drugs such as receptor blockers. [0003] At present, optically active 1,2-epoxy and 1,2-diol are mainly obtained by chemical meth...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N9/14C12P41/00
Inventor 李祖义金浩
Owner SHANGHAI INST OF ORGANIC CHEM CHINESE ACAD OF SCI
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