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Microorganism of producing D-pantothenic acid enternal ester hydrolase and process for preparing D-pantothenic acid thereof

A pantolactone and microorganism technology, applied in the field of biocatalysis research, can solve the problems of long fermentation time, low enzyme utilization rate, short enzyme production time, etc.

Active Publication Date: 2006-06-28
重庆鑫富化工有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, Fusarium oxysporum IFO5942 has defects such as long fermentation time (7 days), only one enzyme conversion, and low enzyme utilization (US Patent 5275949, 1994); and Fusarium moniliforme SW-902 (i.e. CGMCC 0536) strain, the fermentation time of the strain is short (2-3 days), the selectivity to D-pantolactone is high, and the optical purity of the product is high (99%e.e) (Chinese patent CN01104070.X, 2001)

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0120] Preliminary screening of strains producing D-pantolactonase

[0121] Against 96 strains of Fusarium, Gibberella, Aspergillus, Penicillium, Rhizopus, Gliocladium and Aureobasidium (Aureobasidium) and other filamentous fungal strains were screened.

[0122]Prepare a medium containing 1% glycerol, 0.5% peptone, 0.5% yeast extract, 0.5% corn steep liquor, pH 7.0, and 1.5% agar, cool to 50°C after sterilization, add 1.5% D-pantolactone and 0.01% bromothymol blue, mix well into the plate. Various filamentous fungi are collected for screening, spread on a plate, and cultured at 25-30° C. for 5-8 days. Observe the discoloration circles around the colonies. The microbial strains that obviously produce yellow discoloration circles are shown in Table 1, mainly including Fusarium, Gibberella, Aspergillus, Penicillium, root Filamentous fungal strains of the genera Rhizopus, Gliocladium, and Aureobasidium. The specific ones with large yellow discoloration circles are: Fusarium mon...

Embodiment 2

[0126] Re-screening of strains producing D-pantolactonase

[0127] Put the above-mentioned Fusarium, Gibberella, Aspergillus, Penicillium, Rhizopus, and Gliocladium that have a discoloration reaction and a large yellow discoloration circle (Gliocladium) and Aureobasidium (Aureobasidium) and other genus strains were inoculated into the sterilized enzyme-producing liquid medium of the following components: glycerol 2%, peptone 1%, yeast extract 0.5%, corn steep liquor 0.2%, pH The value is 6.5, the liquid volume in the triangular flask is 100mL / 500mL, the rotation speed of the shaker flask is 120r / min, the temperature is controlled at 28°C, and the wet mycelium is collected by suction filtration after 5 days of cultivation. Take 1g of wet mycelium, place it in a 500ml Erlenmeyer flask, add 50ml of A solution and 50ml of B solution (A solution: 20% DL-pantolactone solution, B solution: 1mol / L Tris-HCl buffer solution, pH 7.5). React at 150 r / min on a shaker at 30°C for 60 minut...

Embodiment 3～20

[0130] Microbial cultivation and selective enzymatic hydrolysis

[0131] Prepare the enzyme-producing medium as 2% glycerol, 1% peptone, 0.5% yeast extract, 0.2% corn steep liquor, pH 6.5, divide into 100mL / 500mL triangular bottles, heat sterilize in a sterilizing pot for 20 minutes, The temperature was 121°C. The bacterial strain Fusarium moniliforme AS3.349, F.moniliforme AS3.3489, F.moniliforme SW-05H, Fusarium oxysporum F. oxysporum AS3.1785, F.oxysporum AS3.1786, F.oxysporum AF93327, F.dimerum AS 3.1831, F.dimerum Penz.AF 93237, F.nygamai AS 3.713, F.proliferatum var.minusAS 3.4726, F.proliferatum var .proliferatum AS 3.4738, F.redolensAS 3.2626, Fusarium solani F.solani var.petroliphilum AS3.4489, Fujikura G.fujikuroi AS3.1756, G.fujikuroi AS3.4663, Fujikura G. .fujikuroi var.moniliformis AS3.4746, Fujikura gibberella Fujikura varietal G.fujikuroi var.fujikuroi AS3.4748 and other strains were inoculated into the culture medium and cultured in shake flasks at 28°C for 5...

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PUM

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Abstract

The invention relates to a method used microorganism enzyme resolution DL-pantoic acid lactone to produce D-pantoic acid. It uses the D-pantoic acid lactone hydrolase strain of the fusarium, gibberella, aspergillus, penicillium, rhizopus, gliocladium, aureobasidium to ferment and culture, uses wet thallus as coarse enzyme, DL-pantoic acid lactone as substrate to produce D-pantoic acid. L- pantoic acid lactone can be reclaimed. The DL-pantoic acid lactone gained by racemization reaction can newly be used to do resolution.

Description

technical field [0001] The invention belongs to the field of biocatalysis research, and relates to a microorganism producing D-pantoate lactone hydrolase and a method for preparing D-pantoate. Background technique [0002] D-pantothenic acid (Pantothenic acid) is an important chiral intermediate for the manufacture of D-pantothenic acid. D-pantothenic acid is a vitamin B group substance and a component of coenzyme A. It participates in the metabolism of sugar, fat and protein. Its products are mostly D- It exists as calcium pantothenate. Calcium D-pantothenate is widely used in medicine, food and feed industry. [0003] At present, D-pantothenic acid is generally prepared by chemical or biological resolution methods. Traditional chemical resolution methods have disadvantages such as high production cost, small production volume, and poor optical purity of products. Among the biological resolution methods, the separation of DL-pantolactone by microbial enzymatic method has...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/18C12P7/40C12P13/02C12R1/66C12R1/77C12R1/80C12R1/845
Inventor 孙志浩过鑫富
Owner 重庆鑫富化工有限公司
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