CN101555460A - Bacillus licheniformis and application thereof - Google Patents
Bacillus licheniformis and application thereof Download PDFInfo
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- CN101555460A CN101555460A CNA2009100451109A CN200910045110A CN101555460A CN 101555460 A CN101555460 A CN 101555460A CN A2009100451109 A CNA2009100451109 A CN A2009100451109A CN 200910045110 A CN200910045110 A CN 200910045110A CN 101555460 A CN101555460 A CN 101555460A
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- bacillus licheniformis
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Abstract
The invention provides a Bacillus licheniformis and a method of using the Bacillus licheniformis to catalyse and deoxidize aromatic nitro compounds into corresponding amine compounds. The microorganism related to the invention is Bacillus licheniformis SPRI-60421 (the preservation registration number is CGMCCNo.2280). Microbial strains with the nitro reduction ability are inoculated into a culture medium for the fermentation culture. After the culturing, the fermentation liquid and the strains are separated. The obtained strains precipitation is added into a transformation liquid containing the aromatic nitro compounds. Thus, the aromatic nitro compounds are transformed into the aromatic amine compounds. After the transformation is finished, somatic cells are removed. After the transformation liquid is extracted via organic solvent, the aromatic nitro compounds which are not transformed completely and the product, namely aromatic amines are separated by conventional separating means of chromatography, distillation, etc.
Description
Technical field
The invention belongs to biological chemical field, specifically, relate to a kind of Bacillus licheniformis, and to use this genus bacillus catalytic reduction aromatic nitro compound be the method for corresponding aminated compounds.
Background technology
Because domestic geographical environment, the weather condition of China vary, what Microbial resources were suitable enriches.People seek new microbial strains by the region at various different latitude, height above sea level and weather.Usually, morphological specificity, cultural characteristic and the physiological and biochemical property etc. by bacterial strain also usually adopt the sequence of the 16srDNA that detects microbial strains to come the clearly classification position of this bacterial classification to the microbial strains discriminating of classifying simultaneously.
By cultivating to gathering the microbial strains of getting throughout the country, in the meta-bolites that obtains, seek biological enzyme, come the various organic unit processes of catalysis, hydration reaction that can the catalysis nitrile compounds as Ntn hydrolase prepares corresponding amide class and organic acid compound, hydrolysis reaction that can the catalysis ester compound as lytic enzyme prepares corresponding organic acid and alcohol compound, or the like.At present, utilize microbial fermentation to cultivate the biological enzyme that obtains and come catalysis to synthesize all kinds of Chemicals, become one of important means of producing large fine chemistry product.
Preparing aromatic amine by aromatic nitro compound is an important organic synthesis unit, is the important channel of preparation arylamine.The product arylamine is widely used in fields such as medicine, agricultural chemicals, dyestuff, auxiliary agent, is important organic synthesis intermediate.Traditional technology for preparing arylamine with aromatic nitro compound mainly contains: in ionogen with iron powder reducing, sodium sulfide reducing, catalytic hydrogenating reduction, electrochemical reduction etc.But all there is deficiency separately in these methods, as: the labour intensity of iron powder reducing method is big, seriously polluted; Sulfuration alkaline process yield is low, wastewater flow rate is big; Shortening method cost height, security are not high; Electrochemical reduction method energy consumption is big, cost is high.
Summary of the invention
The present inventor gathers and is separated to a strain bacterial strain in the geographic soil in Yancheng, China Jiangsu Province, be numbered SPRI-60421, by mensuration and analyses such as cultural characteristic, biochemical character, identify that it is Bacillus licheniformis (Bacillus licheniformis) to its 16SrDNA sequence; Discover that further the fermenting culture of this bacterial strain has the nitroreduction ability, aromatic nitro compound can be catalysed and reduced into the corresponding aroma aminated compounds.
Therefore, primary and foremost purpose of the present invention is just providing a kind of Bacillus licheniformis SPRI-60421 and cultural method thereof.
Another object of the present invention is to provide a kind of SPRI-60421 catalytic reduction to prepare the application of aromatic amine compounds.
Bacillus licheniformis of the present invention (Bacillus licheniformis) SPRI-60421 submits Chinese common micro-organisms preservation center preservation on December 05th, 2007, and preserving number is CGMCCNo.2280.
After measured, the 16SrDNA sequence of this bacterial strain is shown in SEQ ID NO:1.
Bacillus licheniformis of the present invention (Bacillus licheniformis) SPRI-60421 is used for catalytic reduction and prepares aromatic amine compounds after fermentation culture.
According to the present invention, the fermentation culture conditions of described Bacillus licheniformis (Bacillus licheniformis) SPRI-60421 was as follows: 37 ℃ of following 230rpm shaking culture 24 hours; The percent weight in volume of substratum is formed and is comprised: 1% glucose, 0.5% yeast extract paste, 0.5% meat extract, 1% sodium-chlor; The PH scope can be 6-8; Culture temperature is at 24-45 ℃, and air flow is with between the ventilation ratio 1: 0.1 to 1 to 1.
According to the present invention, described Bacillus licheniformis (Bacillus licheniformis) SPRI-60421 can not only be applied to the single nitroaromatic of catalysis efficiently, and the dinitrobenzene aromatic compound that can also be applied to reduce efficiently simultaneously is corresponding amine or two amine products.Its catalyzed reaction can be represented with following reaction expression:
According to the present invention, described Bacillus licheniformis (Bacillus licheniformis) SPRI-60421 separates fermented liquid after cultivating and finishing with thalline, the bacterial sediment that obtains joins in the conversion fluid that contains aromatic nitro compound, makes aromatic nitro compound be converted into aromatic amine compounds.
According to the present invention, after transform finishing, remove somatic cells, conversion fluid with organic solvent extraction after, the aromatic nitro compound that will not transform fully with conventional separation means such as chromatography, rectifying separates with the product aromatic amine.
Embodiment
Below by specific embodiment, further specify the present invention.
Bacillus licheniformis of the present invention (Bacillus licheniformis) SPRI-60421 submits Chinese common micro-organisms preservation center preservation on December 05th, 2007, and preserving number is CGMCCNo.2280.
The related starting material of following examples all can obtain by commercially available, and the concentration that relates to is the bulking value specific concentration.
Sequence and the bacterium classification result of morphological specificity, cultural characteristic and the 16srDNA of Bacillus licheniformis of the present invention (Bacillus licheniformis) SPRI-60421 are as follows:
1, morphological specificity and cultural characteristic:
Bacterial strain produces gemma in process of growth.This bacterium thalline is shaft-like, cultivates generation gemma in back about 36h, gives birth in the gemma.On the nutrient agar plate, the initial stage bacterium colony is smooth shape, and it is aqueous that the later stage is row, edge hair fold.According to " the outstanding Bacteria Identification handbook of uncle and " common bacteria system identification handbook " method are carried out biochemical tests such as salt tolerance, thermotolerance, methyl red, its cultural characteristic such as table 1:
Table 1, cultural characteristic
2, the 16s rDNA sequence of bacterial strain SPRI-60421 of the present invention is shown in SEQ ID NO:1.
The 16SrDNA sequential analysis: extract genomic dna with the bacteriolyze enzyme process from new fresh thalli, adopt universal primer to carry out the 16SrDNA amplification, the PCR product after testing, directly use Taq Deoxy behind the purifying
Terminator Cyele Sequencing Ki t order-checking, electrophoresis and number Applied Biosystems DNASequencer (model377) carry out automatically.
According to form and cultural characteristic and 16s rDNA sequential analysis, judge that this bacterial strain belongs to Bacillus licheniformis (Bacillus licheniformis).
The cultural method of this described Bacillus licheniformis SPRI-60421 of invention can be conventional method for cultivation of bacteria.In order to help payable cultivation, above-mentioned production bacterium should be carried out the deep-layer liquid aerated culture.
Nutrition source in the substratum be there is no special regulation, can make and contain carbon source, nitrogenous source and other nutrition sources that is usually used in microbial culture in the substratum.Wherein carbon source can be starch, dextrin, glucose, glycerine, sucrose, N.F,USP MANNITOL etc., and nitrogenous source can be meat peptone, soybean cake powder, peanut powder, yeast, meat extract, rice bran, corn steep liquor, wheat skin, ammonium salt, urea, nitrate and other organic or inorganic nitrogenous compounds.Other nutrition sources can suitably add some inorganic salts, for example metal-salts such as salt, phosphoric acid salt and potassium, calcium, manganese, iron.Can add in case of necessity that some move, plant, the mineral wet goods is as defoamer.
Culture condition such as temperature, PH, time be there is no strict restriction, be as the criterion with the production that is suitable for producing bacterium, and to select to produce the highest condition of nitroreductase for well.For example, the PH scope of substratum can be 6-8, with near neutral for well; Culture temperature is at 24-45 ℃, and air flow is with (ventilation ratio: refer to that per minute passes through the volume of air of nutrient solution and the ratio of nutrient solution volume) between the ventilation ratio 1: 0.1 to 1 to 1.The component of these substratum, PH, culture temperature, aeration condition etc. all should carry out suitable adjustment according to practical situation, to obtain best effect.
The microbial culture medium that above-mentioned fermentation culture is good separates with thalline, can adopt the centrifugal separation method greater than 1000rpm, or filter separation method obtains thalline.
A kind of application of Bacillus licheniformis SPRI-60421 of the present invention, the biocatalysis reduction reaction that can be used for.Can in water or water-organic biphasic system, carry out.The water conversion condition is: concentration of substrate is smaller or equal to 0.2g/L, in the neutrality of any one PH6.5-9.5 in the damping fluid of meta-alkalescence.Organic phase is N, dinethylformamide, methylene dichloride, N-Methyl pyrrolidone, dimethyl sulfoxide (DMSO) or the less organic solvent of other pair cell toxicity.Water-organic phase biphasic system is the mixture of above-mentioned water and organic phase.The invert point scope is at 20-40 ℃, and 50-300rpm shakes bottle vibration ventilation or stirs ventilation or pipeline ventilation oxygen-supplying, reaction times 1-48 hour.After transforming end, conversion fluid and thalline are separated by centrifugal or filtration, can obtain to contain the conversion fluid of aromatic amine.
Need to add concentration in the above-mentioned conversion fluid greater than 0.01% carbon source material, to keep thalline basal metabolism vigor.Carbon source can be other any saccharics such as glucose, sucrose, fructose.
After biocatalysis reduction reaction of the present invention finishes, can in above-mentioned conversion fluid, add organic solvents such as methylene dichloride, trichloromethane, tetrachloromethane, ethyl acetate, N-Methyl pyrrolidone, vibration, extraction, required product and unconverted substrate completely enter in the organic phase.After the static or centrifugal layering, organic phase is removed organic solvent through concentrating under reduced pressure, can obtain the thick product of aromatic amine.This product can obtain the pure product of aromatic amine through means such as chromatography, rectifying.
The fermentation culture of embodiment 1, Bacillus licheniformis SPRI-60421
The LB substratum (biochemical technology handbook---substratum commonly used) of 200mL is put into the 500mL triangular flask, 121 ℃ of high pressure steam sterilizations, insert Bacillus licheniformis of the present invention (Bacillus licheniformis) SPRI-60421CGMCC No.2280 behind the cool to room temperature, after cultivating 24 hours under 37 ℃ and the 230rpm vibration, the centrifugation thalline.
The fermentation culture of embodiment 2, Bacillus licheniformis SPRI-60421
The substratum that 200ml is contained 1% glucose, 0.5% yeast extract paste, 0.5% meat extract, 1% sodium-chlor is put into the 500ml triangular flask, 121 ℃ of high pressure steam sterilizations, insert Bacillus licheniformis of the present invention (Bsciilus licheniformis) SPRI-60421CGMCC No.2280 behind the cool to room temperature, after cultivating 24 hours under 37 ℃ and the 230rpm vibration, the centrifugation thalline.
The biocatalysis reduction reaction of embodiment 3, Meta-dinitrobenzene
The thalline of getting the 100mL bacterium liquid of embodiment 1 gained joins in the 250mL triangular flask of the 50mL phosphoric acid buffer (PH8.0) that contains 0.03% Meta-dinitrobenzene and 0.5% glucose, the ventilation of vibrating of 20 ℃, 300rpm, carry out Meta-dinitrobenzene conversion of resting cells reaction (annotate: conversion of resting cells reaction definition is meant that the bacterial sediment that will obtain after the fermented liquid centrifugation joins the catalytic reduction reaction that carries out in the conversion fluid that contains aromatic nitro compound, down with.), after 24 hours, stop conversion reaction.Adopt the TLC thin layer to carry out trace analysis in the reaction process.
The centrifugal thalline of removing, twice of 10mL dichloromethane extraction of supernatant liquor.The concentrating under reduced pressure organic phase obtains the mixture of faint yellow pentanoic and m-nitraniline and Meta-dinitrobenzene.Obtain each single component through conventional separation means such as chromatography, rectifying again, through conventional liquid-phase chromatographic analysis, each component concentration is all greater than 98%.
The biocatalysis reduction reaction of embodiment 4, Ortho Nitro Toluene
The thalline of getting the 100mL bacterium liquid of embodiment 2 gained joins in the 250mL triangular flask of the 50mL phosphoric acid buffer (PH8.0) that contains 0.04% Ortho Nitro Toluene and 0.5% glucose, the ventilation of vibrating of 25 ℃, 250rpm, carry out the conversion of resting cells reaction of Ortho Nitro Toluene, after 1 hour, stop conversion reaction.Adopt HPLC or TLC thin layer to carry out trace analysis in the reaction process.
The centrifugal thalline of removing, twice of 10mL dichloromethane extraction of supernatant liquor.The concentrating under reduced pressure organic phase obtains content and is about o-toluidine crude product about 90%.Obtain the pure product of o-toluidine through conventional separation means such as chromatography, rectifying again, through conventional liquid-phase chromatographic analysis, content is greater than 98%.
The biocatalysis reduction reaction of embodiment 5, p-nitrophenyl methane amide
The thalline of getting the 100mL bacterium liquid of embodiment 2 gained joins in the 250mL triangular flask of the 50mL phosphoric acid buffer (PH8.0) that contains 0.06% p-nitrophenyl methane amide and 0.5% sucrose, the ventilation of vibrating of 30 ℃, 200rpm, carry out the conversion of resting cells reaction of p-nitrophenyl methane amide, after 6 hours, stop conversion reaction.Adopt the TLC thin layer to carry out trace analysis in the reaction process.
The centrifugal thalline of removing, twice of 10mL dichloromethane extraction of supernatant liquor.The concentrating under reduced pressure organic phase obtain content be about about 90% to methane amide aniline crude product.Obtain the pure product of methane amide aniline through conventional separation means such as chromatography, rectifying, through conventional liquid-phase chromatographic analysis, content is greater than 98% again.
The biocatalysis reduction reaction of embodiment 6,6-chloro-2-nitrotoluene
The thalline of getting the 100mL bacterium liquid of embodiment 2 gained joins in the 250mL triangular flask of the 50mL phosphoric acid buffer (PH8.0) that contains 0.04%6-chloro-2-nitrotoluene and 0.5% fructose, the ventilation of vibrating of 35 ℃, 150rpm, carry out the conversion of resting cells reaction of 6-chloro-2-nitrotoluene, after 12 hours, stop conversion reaction.Adopt the TLC thin layer to carry out trace analysis in the reaction process.
The centrifugal thalline of removing, twice of 10mL dichloromethane extraction of supernatant liquor.The concentrating under reduced pressure organic phase obtains content and is about 2-methyl-3-chloroaniline crude product about 90%.Obtain 2-methyl-pure product of 3-chloroaniline through conventional separation means such as chromatography, rectifying again, through conventional liquid-phase chromatographic analysis, content is greater than 98%.
Embodiment 7,4-chloro-2, the biocatalysis reduction reaction of 5-dimethoxy oil of mirbane
The thalline of getting the 100mL bacterium liquid of embodiment 2 gained joins and contains 0.04%4-chloro-2, in the 250mL triangular flask of the 50mL phosphoric acid buffer (PH8.0) of 5-dimethoxy oil of mirbane and 0.5% glucose, 40 ℃ are stirred uncovered ventilation, carry out 4-chloro-2, the conversion of resting cells reaction of 5-dimethoxy oil of mirbane, after 20 hours, stop conversion reaction.Adopt the TLC thin layer to carry out trace analysis in the reaction process.
The centrifugal thalline of removing, twice of 10mL dichloromethane extraction of supernatant liquor.The concentrating under reduced pressure organic phase obtains content and is about 4-chloro-2 about 90%, 5-dimethoxyaniline crude product.Obtain 4-chloro-2 through conventional separation means such as chromatography, rectifying again, the pure product of 5-dimethoxyaniline, through conventional liquid-phase chromatographic analysis, content is greater than 98%.
The two-phase biocatalysis reduction reaction of embodiment 8, p-nitrophenyl methane amide
The thalline of getting the 100mL bacterium liquid of embodiment 2 gained joins in the 250mL triangular flask of the 50mL phosphoric acid buffer (PH8.0) that contains 0.5% p-nitrophenyl methane amide and 0.5% sucrose, in aforesaid conversion fluid, add dimethyl sulfoxide (DMSO) (DMSO) then, every 100ml conversion fluid adds the 10ml dimethyl sulfoxide (DMSO), 30 ℃ of pipeline ventilations, carry out the conversion of resting cells reaction of p-nitrophenyl methane amide, after 30 hours, stop conversion reaction.Adopt the TLC thin layer to carry out trace analysis in the reaction process.
The centrifugal thalline of removing, twice of 10mL dichloromethane extraction of supernatant liquor.The concentrating under reduced pressure organic phase obtain content be about about 90% to methane amide aniline crude product.Obtain the pure product of methane amide aniline through conventional separation means such as chromatography, rectifying, through conventional liquid-phase chromatographic analysis, content is greater than 98% again.
The two-phase biocatalysis reduction reaction of embodiment 9, Meta-dinitrobenzene
The thalline of getting the 100mL bacterium liquid of embodiment 2 gained joins in the 250mL triangular flask of the 50mL phosphoric acid buffer (PH8.0) that contains 0.5% Meta-dinitrobenzene and 0.5% fructose, in aforesaid conversion fluid, add methylene dichloride then, every 100ml conversion fluid adds 10mlN, dinethylformamide, 20 ℃ are stirred uncovered ventilation, carry out the conversion of resting cells reaction of Meta-dinitrobenzene, after 36 hours, stop conversion reaction.Adopt the TLC thin layer to carry out trace analysis in the reaction process.
The centrifugal thalline of removing, twice of 10mL dichloromethane extraction of supernatant liquor.The concentrating under reduced pressure organic phase obtain content be about about 90% between the pentanoic crude product.Obtain the pure product of a pentanoic through conventional separation means such as chromatography, rectifying again, through conventional liquid-phase chromatographic analysis, content is greater than 98%.
The two-phase biocatalysis reduction reaction of embodiment 10, Ortho Nitro Toluene
The thalline of getting the 100mL bacterium liquid of embodiment 2 gained joins in the 250mL triangular flask of the 50mL phosphoric acid buffer (PH8.0) that contains 0.5% Ortho Nitro Toluene and 0.5% glucose, in aforesaid conversion fluid, add N-Methyl pyrrolidone then, every 100ml conversion fluid adds the 10ml N-Methyl pyrrolidone, 40 ℃ of pipeline ventilations, carry out the conversion of resting cells reaction of Ortho Nitro Toluene, after 48 hours, stop conversion reaction.Adopt the TLC thin layer to carry out trace analysis in the reaction process.
The centrifugal thalline of removing, twice of 10mL dichloromethane extraction of supernatant liquor.The concentrating under reduced pressure organic phase obtains content and is about o-toluidine crude product about 90%.Obtain the pure product of o-toluidine through conventional separation means such as chromatography, rectifying again, content is greater than 98%.
In sum, the present invention is the biocatalysis process of carrying out the catalytic reduction aromatic nitro compound with a kind of Bacillus licheniformis SPRI-60421, compare with technology such as traditional iron powder reducing, sodium sulfide reducing, catalytic hydrogenating reduction, electrochemical reductions, have characteristics such as catalytic condition gentleness, Environmental compatibility is good, the single-minded selectivity of catalysis is good, provide a kind of new approach for prepare aromatic amine by aromatic nitro compound.
Sequence table
＜110〉Shanghai Pesticide Research Institute Nanjing Normal University
＜120〉a kind of Bacillus licheniformis and application thereof
<130>P5081086
<160>1
<170>PatentIn?version?3.3
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<211>1425
<212>DNA
<213>Bacillus?licheniformis
<220>
<221>16srDNA
<222>(1)..(1425)
<400>1
cggcggctgg?ctccaaaagg?ttacctcacc?gacttcgggt?gttacaaact?ctcgtggtgt 60
gacgggcggt?gtgtacaagg?cccgggaacg?tattcaccgc?ggcatgctga?tccgcgatta 120
ctagcgattc?cagcttcacg?cagtcgagtt?gcagactgcg?atccgaactg?agaacagatt 180
tgtgggattg?gcttagcctc?gcggcttcgc?tgccctttgt?tctgcccatt?gtagcacgtg 240
tgtagcccag?gtcataaggg?gcatgatgat?ttgacgtcat?ccccaccttc?ctccggtttg 300
tcaccggcag?tcaccttaga?gtgcccaact?gaatgctggc?aactaagatc?aagggttgcg 360
ctcgttgcgg?gacttaaccc?aacatctcac?gacacgagct?gacgacaacc?atgcaccacc 420
tgtcactctg?cccccgaagg?ggaagcccta?tctctagggt?tgtcagagga?tgtcaagacc 480
tggtaaggtt?cttcgcgttg?cttcgaatta?aaccacatgc?tccaccgctt?gtgcgggccc 540
ccgtcaattc?ctttgagttt?cagtcttgcg?accgtactcc?ccaggcggag?tgcttaatgc 600
gtttgctgca?gcactaaagg?gcggaaaccc?tctaacactt?agcactcatc?gtttacggcg 660
tggactacca?gggtatctaa?tcctgttcgc?tccccacgct?ttcgcgcctc?agcgtcagtt 720
acagaccaga?gagtcgcctt?cgccactggt?gttcctccac?atctctacgc?atttcaccgc 780
tacacgtgga?attccactct?cctcttctgc?actcaagttc?cccagtttcc?aatgaccctc 840
cccggttgag?ccgggggctt?tcacatcaga?cttaagaaac?cgcctgcgcg?cgctttacgc 900
ccaataattc?cggacaacgc?ttgccaccta?cgtattaccg?cggctgctgg?cacgtagtta 960
gccgtggctt?tctggttagg?taccgtcaag?gtgccgccct?attcgaacgg?tacttgttct 1020
tccctaacaa?cagagtttta?cgatccgaaa?accttcatca?ctcacgcggc?gttgctccgt 1080
cagactttcg?tccattgcgg?aagattccct?actgctgcct?cccgtaggag?tctgggccgt 1140
gtctcagtcc?cagtgtggcc?gatcaccctc?tcaggtcggc?tacgcatcgt?cgccttggtg 1200
agccgttacc?tcaccaacta?gctaatgcgc?cgcgggtcca?tctgtaagtg?gtagctaaaa 1260
gccacctttt?atgattgaac?catgcggttc?aatcaagcat?ccggtattag?ccccggtttc 1320
ccggagttat?cccagtctta?caggcaggtt?acccacgtgt?tactcacccg?tccgccgctg 1380
acctaaggga?gcaagctccc?gtcggtccgc?tcgacttgca?tgtat 1425
Claims (12)
1, a kind of Bacillus licheniformis (Bacillus licheniformis) SPRI-60421 is characterized in that the preserving number of this bacterial strain is CGMCC No.2280.
2, Bacillus licheniformis SPRI-60421 as claimed in claim 1 is characterized in that, this bacterial strain has the 16SrDNA sequence shown in the SEQ ID NO:1.
3, Bacillus licheniformis SPRI-60421 as claimed in claim 1 is characterized in that, the fermentation culture conditions of this bacterial strain is: 37 ℃ of following 230rpm shaking culture 24 hours; The percent weight in volume of substratum is formed and is comprised: 1% glucose, 0.5% yeast extract paste, 0.5% meat extract, 1% sodium-chlor; PH6-8; Culture temperature is at 24-45 ℃, and air flow is for ventilating than between (1: 0.1)～(1: 1).
4, as the application of each described Bacillus licheniformis SPRI-60421 of claim 1-3, it is characterized in that, be used for the aromatic nitro compound catalytic reduction is prepared the aromatic amine compounds.
5, application as claimed in claim 4, it is characterized in that, described catalytic reduction is that the bacterial sediment that described Bacillus licheniformis SPRI-60421 fermentation back is obtained is joined in the conversion fluid that contains aromatic nitro compound, makes aromatic nitro compound be converted into aromatic amine compounds.
As claim 4 or 5 described application, it is characterized in that 6, described catalytic reduction reaction carries out in water or water-organic biphasic system.
7, application as claimed in claim 6 is characterized in that, the conversion condition in the described water system is: concentration of substrate carries out in the damping fluid of meta-alkalescence in the neutrality of any one PH6.5-9.5 smaller or equal to 0.2g/L.
8, application as claimed in claim 6 is characterized in that, described water-organic biphasic system is water and N, the mixture of the organic solvent that dinethylformamide, dimethyl sulfoxide (DMSO), N-Methyl pyrrolidone, methylene dichloride or other pair cell toxicity are less.
9, as claim 4 or 5 described application, it is characterized in that described catalytic reaction condition is: invert point is at 20-40 ℃, and 50-300rpm shakes bottle vibration ventilation, or stirs ventilation or pipeline ventilation oxygen-supplying, reaction times 1-48 hour.
As claim 4 or 5 described application, it is characterized in that 10, add concentration in the conversion fluid of described catalytic reduction reaction greater than 0.01% carbon source material, carbon source is glucose, sucrose or fructose.
As claim 4 or 5 described application, it is characterized in that 11, being used for single nitroaromatic catalytic reduction is a corresponding amine product.
As claim 4 or 5 described application, it is characterized in that 12, being used for dinitrobenzene aromatic compound catalytic reduction is corresponding amine or two amine products.
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| CNA2009100451109A CN101555460A (en) | 2009-01-09 | 2009-01-09 | Bacillus licheniformis and application thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106947795A (en) * | 2017-05-13 | 2017-07-14 | 太原理工大学 | A kind of method that living things catalysis synthesizes 2 thiophene ethamines |
| CN108192933A (en) * | 2018-01-23 | 2018-06-22 | 上海韶屹生物科技有限公司 | A kind of method that enzyme process prepares aliskiren key intermediate |
| CN113817075A (en) * | 2021-09-24 | 2021-12-21 | 郑州大学 | Bacillus licheniformis extracellular polymer metal iron complex and preparation method and application thereof |
-
2009
- 2009-01-09 CN CNA2009100451109A patent/CN101555460A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106947795A (en) * | 2017-05-13 | 2017-07-14 | 太原理工大学 | A kind of method that living things catalysis synthesizes 2 thiophene ethamines |
| CN106947795B (en) * | 2017-05-13 | 2020-05-22 | 太原理工大学 | Method for synthesizing 2-thiophene ethylamine through biocatalysis |
| CN108192933A (en) * | 2018-01-23 | 2018-06-22 | 上海韶屹生物科技有限公司 | A kind of method that enzyme process prepares aliskiren key intermediate |
| CN113817075A (en) * | 2021-09-24 | 2021-12-21 | 郑州大学 | Bacillus licheniformis extracellular polymer metal iron complex and preparation method and application thereof |
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