CN109609389B - Penicillium oxalicum ZJB16086 and application thereof in synthesis of R-2- (4-hydroxyphenoxy) propionic acid - Google Patents
Penicillium oxalicum ZJB16086 and application thereof in synthesis of R-2- (4-hydroxyphenoxy) propionic acid Download PDFInfo
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Abstract
The invention discloses a Penicillium oxalicum (Penicillium oxalicum) ZJB16086 and application thereof in synthesizing R-2- (4-hydroxyphenoxy) propionic acid, wherein the preservation number is CCTCC NO: M2018409. The invention separates penicillium oxalicum (Penicillium oxalicum) CCTCC NO: M2018409 which effectively converts R-2-phenoxypropionic acid to synthesize R-2- (4-hydroxyphenoxy) propionic acid from humus soil, and can synthesize 2.6g/L of R-2- (4-hydroxyphenoxy) propionic acid under the condition of 10g/L of substrate, wherein the conversion rate is 26%.
Description
Technical Field
A microorganism for synthesizing R-2- (4-hydroxyphenoxy) propionic acid by converting R-2 phenoxypropionic acid and a screening method thereof belong to the technical field of bioengineering, in particular to penicillium oxalicum for synthesizing R-2- (4-hydroxyphenoxy) propionic acid by converting R-2 phenoxypropionic acid.
Background
R-2- (4-hydroxyphenoxy) propionic acid is a key intermediate in the synthesis process of phenoxy carboxylic acid and aryloxy phenoxy propionic acid herbicides, and can be used for synthesizing herbicides such as fenoxaprop (soybean, rice), fluazifop-p-butyl (rape, potato), clodinafop-propargyl (grain) and the like. As phenoxy carboxylic acid herbicides are receiving increased attention, their advantages as herbicides are also increasingly recognized. As chiral pesticide with enantiomer, compared with racemic pesticide, the chiral pesticide has low efficiency and high environmental pollution, and the optically pure phenoxy carboxylic acid herbicide has the advantages of high weeding efficiency, low harm to crops, low environmental pollution and the like. The synthesis of phenoxy carboxylic acid herbicides uses chiral R-2- (4-hydroxyphenoxy) propionic acid as an intermediate, so the development of efficient synthesis methods of R-2- (4-hydroxyphenoxy) propionic acid is attracting much attention.
Elango V et al (U.S. Pat. No. 3, 5008439A) reported that p-hydroxyacetophenone as a starting material was reacted with an α -halopropionate, oxidized and hydrolyzed to give a racemic modification of 2- (4-hydroxyphenoxy) propionic acid, which was further resolved to give R-2- (4-hydroxyphenoxy) propionic acid of high optical purity. The technical route is systematically improved by Shenhengxi et al (synthetic chemistry, 2006, 04: 398-. Although the product is easy to separate and purify, the method needs multi-step reaction to complete, so that the yield of the product is not high, and meanwhile, reaction raw materials and reagents are not suitable for large-scale production.
Umbelliferae et al (proceedings of Zhejiang university of Industrial science 2009, 04: 362-365) reported that high-optical-purity R-2- (4-hydroxyphenoxy) propionic acid was produced by esterification, sulfonylation, etherification and the like using L-lactic acid as a starting compound. Wherein the configuration is reversed in the reaction of S-2- (4-tosyloxy) propionic acid to generate R-2- (4-hydroxyphenoxy) propionic acid, and the reaction mechanism is as follows: under the condition of sodium hydroxide, hydroquinone is firstly generated into sodium phenolate, ethyl S-2- (4-tosyloxy) propionate is changed into sodium salt, sodium ions are positively charged, oxygen of the sodium phenolate is negatively charged, chiral carbon in the sodium S-2- (4-tosyloxy) propionate is attacked from the surface with small steric hindrance, and after an intermediate is formed, the configuration is inverted and is changed into an R type. Wuchunlei et al (Guangzhou chemical 2015, 08:92-93) reported that L-alanine was used as a starting compound, and the product R-2- (4-hydroxyphenoxy) propionic acid was produced by steps of chlorination, etherification and acidification, in which the configuration was inverted during the etherification reaction. The process route of the L-alanine and the L-lactic acid is simpler, the reaction steps are fewer, but the conditions are difficult to control. Because hydroquinone participates in the reaction process, the hydroquinone is easy to be oxidized to generate quinone in an alkaline environment, so that the disubstituted product of the hydroquinone and the hydroquinone which are not completely reacted is difficult to remove in the later purification treatment, and the purity of the product is influenced. Therefore, in the synthesis process, the reaction conditions and the material ratio need to be strictly controlled to prevent side reactions from occurring.
At present, few studies have been made in foreign countries on the biological production of R-2- (4-hydroxyphenoxy) propionic acid, and among them, BASF company Dingler et al (Pest Management Science,2015,46(1):33-35) have reported that R-2- (4-hydroxyphenoxy) propionic acid is synthesized by hydroxylating the substrate R-2-phenoxypropionic acid by Beauveria bassiana. Kinne M et al (Tetrahedron Letters,2008,49(41):5950-2O2Under the condition, the racemic 2-phenoxypropionic acid is selectively hydroxylated to generate R-2- (4-hydroxyphenoxy) propionic acid. With the attention of people on environmental protection, the original chemical industry is impacted to a certain extent, and meanwhile, the rapid development of the green biological manufacturing industry is promoted, so that the biological method for preparing the R-2- (4-hydroxyphenoxy) propionic acid has a great application prospect.
Disclosure of Invention
The invention aims to provide penicillium oxalicum (Penicillium oxalicum) ZJB16086 and application thereof in the synthesis of R-2- (4-hydroxyphenoxy) propionic acid by converting R-2 phenoxypropionic acid.
The technical scheme adopted by the invention is as follows:
the invention provides penicillium oxalicum (Penicillium oxalicum) ZJB16086 for synthesizing R-2- (4-hydroxyphenoxy) propionic acid by converting R-2 phenoxypropionic acid, which is preserved in China center for type culture Collection with the preservation number of CCTCC NO: M2018409, the preservation date of 2018, 6 and 28 days, the preservation address: wuhan university, Wuhan, China, zip code 430072.
The penicillium oxalicum CCTCC NO: M2018409 is obtained by performing enrichment culture twice on humus soil, selecting a strain which can grow in a culture medium containing a substrate R-2-phenoxypropionic acid, inoculating the strain into a 96-pore plate fermentation primary sieve, screening a primary sieve strain which can convert the substrate R-2-phenoxypropionic acid into R-2- (4-hydroxyphenoxy) propionic acid by adopting a sodium nitrite color development method, and then transferring the primary sieve strain into a shake flask containing a fermentation culture medium for fermentation and re-screening.
The invention also provides application of the penicillium oxalicum ZJB16086 in catalyzing R-2 phenoxypropionic acid to synthesize R-2- (4-hydroxyphenoxy) propionic acid, wherein fermentation liquor obtained by fermentation culture of the penicillium oxalicum ZJB16086 is used as a catalyst and a reaction medium, a substrate R-2 phenoxypropionic acid is added, conversion reaction is carried out at the temperature of 25-30 ℃ and the shaking table rotation speed of 150-. The final concentration of the substrate in the fermentation liquor is 10-50g/L, preferably 10 g/L; the content of wet bacteria in the fermentation liquor is 15-20 g/L, and preferably 15 g/L.
Further, the fermentation broth obtained by fermentation culture of the penicillium oxalicum ZJB16086 is prepared by the following method: inoculating penicillium oxalicum ZJB16086 to a fermentation conversion culture medium, and performing fermentation culture (preferably culturing until wet thallus in the fermentation broth reaches 15-20 g/L) at the temperature of 25-30 ℃ and the shaking table rotating speed of 150-; the fermentation transformation medium comprises the following components: 5-20g/L glucose, 5-10g/L yeast extract, 5-10g/L ammonium sulfate, 0.5-1.0g/L magnesium sulfate heptahydrate, 0.05-0.12g/L manganese sulfate monohydrate, 1.5-2.5g/L potassium dihydrogen phosphate, 3.6-6.0g/L dipotassium hydrogen phosphate trihydrate, 1-5mL/L trace element liquid, deionized water as a solvent, 5M sodium hydroxide solution for adjusting the pH value to 6.8, and sterilizing at 115 ℃ for 20 min; the trace element liquid: 2g/L of ferrous sulfate heptahydrate, 100mg/L of zinc sulfate (II) tetrahydrate, 300mg/L of boric acid, 200mg/L of cobalt chloride (II) hexahydrate, 10mg/L of copper chloride (II) dihydrate, 20mg/L of nickel chloride (II) hexahydrate and 30mg/L of sodium molybdate dihydrate, and deionized water is used as a solvent.
Further, the fermentation medium composition is: 5g/L of glucose, 5g/L of yeast extract, 5g/L of ammonium sulfate, 0.5g/L of magnesium sulfate heptahydrate, 0.05g/L of manganese sulfate monohydrate, 1.5g/L of potassium dihydrogen phosphate, 3.6g/L of dipotassium hydrogen phosphate trihydrate, 1mL/L of trace element liquid, deionized water as a solvent, and 5M sodium hydroxide solution is used for adjusting the pH value to be 6.8.
Further, before the penicillium oxalicum ZJB16086 is inoculated to the fermentation transformation medium, slant culture and seed culture are carried out, specifically: inoculating penicillium oxalicum ZJB16086 to a slant culture medium, and performing static culture at 28 deg.C for 6-7d to obtain slant strain; selecting thalli from the inclined plane, inoculating the thalli into a 250mL shaking flask filled with 50mL seed culture medium, and culturing at 28 ℃ and 150-; inoculating the seed solution into a 250mL shake flask containing 50mL fermentation transformation culture medium by an inoculation amount with the volume concentration of 3% for fermentation transformation culture; the slant culture medium (PDA medium): 200g/L of potato, 20g/L of glucose, 20g/L of agar and deionized water as a solvent, wherein the pH is natural, and the potato is sterilized at 115 ℃ for 20 min; the seed medium (PDA medium): 200g/L of potato, 20g/L of glucose and deionized water as a solvent, wherein the pH is natural, and the potato is sterilized at 115 ℃ for 20 min.
The invention establishes a method for quickly and effectively screening a microorganism strain for synthesizing R-2- (4-hydroxyphenoxy) propionic acid by a biological method, namely, designs an experimental scheme aiming at screening microorganisms which can grow in a culture medium containing a substrate R-2-phenoxypropionic acid and can convert the R-2-phenoxypropionic acid to synthesize the R-2- (4-hydroxyphenoxy) propionic acid: firstly, enriching microorganisms in humus soil in a culture medium containing a substrate R-2-phenoxypropionic acid, and primarily screening strains capable of growing on an enrichment culture medium plate. Then the ability of converting R-2-phenoxypropionic acid into R-2- (4-hydroxyphenoxy) propionic acid is detected, namely, a single colony which grows out is selected and inoculated into a 96-deep-hole plate containing a fermentation culture medium (the formula is the same as that of an enrichment culture medium) for culture, and the ability of each strain to convert a substrate into a product is detected by a sodium nitrite color development method. And preserving the screened strains to obtain primary screened strains. And respectively inoculating the strains obtained by primary screening into 250mL triangular flasks filled with 50mL fermentation medium (the formula of the enrichment medium is the same as that of the enrichment medium) for secondary screening, culturing for about 7 days, and detecting the concentration of the R-2- (4-hydroxyphenoxy) propionic acid in the fermentation liquor by HPLC. Through the screening work of a large number of strains of a plurality of soil samples, ZJB16086 strains with higher transformation capacity are screened from more than 30 separated soil microorganisms.
The microorganism ZJB16086 which is obtained by screening and can effectively convert R-2-phenoxypropionic acid to synthesize R-2- (4-hydroxyphenoxy) propionic acid is cultured on a solid flat plate for about 7 days, the diameter of a bacterial colony is about 2.0cm, the bacterial colony is loose, the texture is villous, the middle part is convex, and the bacterial colony is green. The back of the colony is light yellow. Observed under an optical microscope, the hyphae are long, dense and branched, and a bunch of conidia is formed at the top of the hyphae, so that the number of spores is large. Observing under electron microscope, the hypha is smooth and branched, and the diameter of the hypha is 2.3-2.5 μm; without apical sac, conidiophores have single stem and are separated, and multiple rounds of branches are like brooms. The conidiophore is oval or approximately spherical, has single spore diameter of 4.0-5.0 μm, and is primarily determined to be Penicillium (Penicillium) according to the microbial morphology analysis. In combination with molecular biological identification, Penicillium oxalicum (Penicillium oxalicum) is considered to belong to the genus Penicillium of the family Eurotiaceae.
Compared with the prior art, the invention has the following beneficial effects: the invention separates penicillium oxalicum (Penicillium oxalicum) CCTCC NO: M2018409 which effectively converts R-2-phenoxypropionic acid to synthesize R-2- (4-hydroxyphenoxy) propionic acid from humus soil, and can synthesize 2.6g/L of R-2- (4-hydroxyphenoxy) propionic acid under the condition of 10g/L of substrate, wherein the conversion rate is 26%.
Drawings
FIG. 1 shows the results of primary screening of the strains, the letters representing the columns and the numbers representing the rows.
FIG. 2 HPLC chromatograms of R-2- (4-hydroxyphenoxy) propionic acid standard (A), R-2-phenoxypropionic acid standard (B) and strain ZJB16086 fermentation conversion solution (C).
FIG. 3 is a front (A) and back (B) morphograms of a ZJB16086 colony.
FIG. 4 shows the mycelium morphology of strain ZJB16086 under an optical microscope at 10X 10 times.
FIG. 5 SEM micrograph of strain ZJB 16086.
FIG. 6 phylogenetic tree of strain ZJB 16086.
The specific implementation mode is as follows:
the following is an example of the screening, identification and fermentative conversion of R-2-phenoxypropionic acid to R-2- (4-hydroxyphenoxy) propionic acid by Penicillium oxalicum (Penicillium oxalicum) CCTCC NO: M2018409.
Example 1: screening of Penicillium oxalicum (Penicillium oxalicum) ZJB16086
(1) Sampling soil from multiple plant growing areas in campus of Zhejiang industry university, weighing 1g of soil sample (wet weight), preparing suspension with 9mL of 0.9% physiological saline, sucking 1mL of the suspension, inoculating the suspension into a 250mL triangular flask containing 50mL of enrichment medium containing 10g/L R-2-phenoxypropionic acid, culturing on a shaking table at 28 ℃ and 150rpm for 1-2d, and performing first enrichment culture; taking 1mL of the bacterial liquid of the first enrichment culture to perform the second enrichment culture in a fresh enrichment culture medium containing 10g/L R-2-phenoxypropionic acid under the same conditions. Carrying out gradient dilution on the bacterial liquid subjected to the second enrichment culture, and respectively taking 100 mu L of diluted 103And 105Spreading the diluted solution on a screening medium plate, and performing static culture in a constant temperature incubator at 28 ℃ for 3-4 d. A single colony growing on the screening plate is picked by a sterile toothpick and inoculated in a 96-deep-well plate (1 mL of fermentation medium containing 10g/L R-2-phenoxypropionic acid is added in each well), after the single colony is cultured for 6-7d at 28 ℃ and 150rpm, the 96-deep-well plate is centrifuged for 15min at 1000rpm, 100 mu L of supernatant is absorbed in the 96-deep-well plate, and the capability of each strain for converting a substrate into a product is detected by a sodium nitrite chromogenic method (figure 1). 64 strains having the transformation ability were selected and deposited.
(2) Inoculating the primary screened strain into a 250mL shaking flask containing 50mL of seed culture medium, culturing at 28 ℃ and 150rpm for 2-3d, inoculating the primary screened strain into a 250mL shaking flask containing 50mL of fermentation culture medium containing 10g/L R-2-phenoxypropionic acid at a volume concentration of 3%, performing fermentation and re-screening at 28 ℃ and 150rpm for 7d, centrifuging at 10000rpm for 10min, filtering the supernatant through a 0.22 μm microporous membrane, and detecting the concentration of R-2-phenoxypropionic acid and R-2- (4-hydroxyphenoxy) propionic acid by HPLC (an Erite C18 reverse phase column of 250mm x 4.6mm, a mobile phase V (a phosphoric acid aqueous solution with pH of 2): V (acetonitrile): 3: 2, a flow rate of 1mL/min, a detector DAD, a detection wavelength of 210nm, and a column temperature of 30 ℃). The retention times of R-2-phenoxypropionic acid and R-2- (4-hydroxyphenoxy) were 8.1min and 3.9min, respectively, as shown in FIG. 2. Screening strains with higher transformation capability to obtain re-screened strains ZJB16086 for slant preservation.
Among them, the enrichment medium of this example consists of: 5g/L of glucose, 5g/L of yeast extract, 5g/L of ammonium sulfate, 0.5g/L of magnesium sulfate heptahydrate, 0.05g/L of manganese sulfate monohydrate, 1.5g/L of potassium dihydrogen phosphate, 3.6g/L of dipotassium hydrogen phosphate trihydrate, 1mL/L of trace element liquid and deionized water as a solvent, adjusting the pH value to 6.8 by using 5M sodium hydroxide solution, and sterilizing at 115 ℃ for 20 min.
The trace element liquid comprises the following components: 2g/L of ferrous sulfate heptahydrate, 100mg/L of zinc sulfate (II) tetrahydrate, 300mg/L of boric acid, 200mg/L of cobalt chloride (II) hexahydrate, 10mg/L of copper chloride (II) dihydrate, 20mg/L of nickel chloride (II) hexahydrate and 30mg/L of sodium molybdate dihydrate, and deionized water is used as a solvent.
Screening medium composition: 20g/L agar is added in the enrichment medium formula.
Slant medium (PDA medium) composition: 200g/L of potato, 20g/L of glucose, 20g/L of agar and deionized water as a solvent, wherein the pH is natural, and the potato is sterilized at 115 ℃ for 20 min.
Seed medium (PDA medium) composition: 200g/L of potato, 20g/L of glucose and deionized water as a solvent, wherein the pH is natural, and the potato is sterilized at 115 ℃ for 20 min.
The fermentation medium is composed of the same enrichment medium.
Sodium nitrite color development method: adding 100 mu L of sample to be detected into a 96 micro-porous plate, adding 100 mu L of 6g/L nitrous acid solution into each hole, reacting for 30min at 60 ℃, observing the color depth, and detecting the light absorption value of the sample at 420nm by using an enzyme-labeling instrument.
Example 2: identification of Strain ZJB16086
(1) Observation of colony morphology
A small amount of spores of the strain are picked by an inoculating needle, a PDA plate is streaked, the strain is cultured for about 7 days at the temperature of 28 ℃, and the single colony morphology is shown in figure 3. The diameter of the bacterial colony is about 2.0cm, the bacterial colony is loose, the texture is villiform, the middle part is convex, and the bacterial colony is green. The back of the colony is light yellow.
(2) Observation of mycelium morphology
Spreading a round filter paper on the bottom of a culture dish with a diameter of 9.0cm, placing a U-shaped glass rod on the round filter paper, placing a clean glass slide and two cover slips on the glass rod, covering the dish with a cover, sterilizing, inoculating, culturing at 28 deg.C for about 7d, and observing the hypha shape under an optical microscope at a power of 10 × 10 times, as shown in FIG. 4. The hyphae are long, dense and branched, and a bunch of conidia is formed at the top of the hyphae, so that the number of the spores is large.
(3) SEM electron microscope observation
The SEM results are shown in FIG. 5. The hypha is smooth and has branches, and the diameter of the hypha is 2.3-2.5 mu m; without apical sac, conidiophores have single stem and are separated, and multiple rounds of branches are like brooms. Conidiophore is oval or nearly spherical, single spore, diameter 4.0-5.0 μm, and according to morphology, the strain is primarily determined to be Penicillium (Penicillium).
(4) Molecular identification
Chromosomal DNA was extracted according to the molecular kit instructions, and 18S rDNA sequences were PCR-amplified using total DNA as template and ITS1 (5'-TCCGTAGGTGAACCTGCGG-3') and ITS4 (5'-TCCTCCGCTTATTGATATGC-3') as forward and reverse primers. The PCR amplification system was (50 mL): 10 × Taq Buffer (Mg)2+)mu.L of 5. mu.L, 1. mu.L of 10mM dNTPs, 0.5. mu.L of Taq DNA polymerase, 0.5. mu.L of each of primers ITS1 and ITS4, 1.5. mu.L of template, ddH2O41. mu.L. The PCR reaction conditions are as follows: pre-denaturation at 94 deg.C for 2min, denaturation at 94 deg.C for 30s, annealing at 50 deg.C for 30s, extension at 72 deg.C for 1min, 35 cycles, preservation at 72 deg.C for 10min, and storage at 4 deg.C. PCR products were assigned to Hangzhou Hingxi Biotechnology Limited for sequencing (SEQ ID NO:1), then searched by BLAST on NCBI website to select rDNA ITS sequences with higher similarity to the sequences, and then generated into a neighbor junction tree (NJ phylogenetic tree) by 1000 repeats using MEGA5 software Align by cluster automatic analysis (FIG. 6). The results show 100% similarity of ZJB16086 to Penicillium oxalicum NRRL 787. Combined with morphological identification and study of the growth characteristics of the strain, it is considered to be a variant of Penicillium oxalicum, named Penicillium oxalicum ZJB16086, which has been deposited in the Chinese dictionaryThe type culture collection center has a collection number of CCTCC NO: M2018409 and a collection date of 2018, 6 months and 28 days.
Any nucleotide sequence obtained by carrying out one or more nucleotide substitutions, deletions or insertions on the nucleotide sequence shown in SEQ NO. 1 is within the protection scope of the present invention as long as the nucleotide sequence has more than 90% of similarity with the nucleotide sequence.
Example 3: penicillium oxalicum CCTCC No. M2018409 fermentation conversion of R-2-phenoxypropionic acid to produce R-2- (4-hydroxyphenoxy) propionic acid
Inoculating penicillium oxalicum CCTCC No. M2018409 to a slant culture medium, and performing static culture at 28 ℃ for 6-7d to obtain slant strains. The thalli is picked from the inclined plane and inoculated into a 250mL shaking flask filled with 50mL seed culture medium, and cultured for 2-3d at 28 ℃ and 200rpm to obtain seed liquid. Inoculating the cultured seed solution into a 250mL shake flask containing 50mL fermentation medium by an inoculation amount with the volume concentration of 3%, culturing at 28 ℃ and 150-200rpm for 2-3d, allowing the wet thallus concentration in the fermentation broth to reach 15g/L, adding R-2 phenoxypropionic acid with the final concentration of 10g/L, and continuing culturing for 6-7 d. The reaction mixture was centrifuged at 10000rpm for 10min, and the supernatant was filtered through a 0.22 μm microfiltration membrane and then subjected to HPLC (Ulipret C18 reverse phase column 250 mm. times.4.6 mm, mobile phase V (aqueous phosphoric acid solution having pH 2): V (acetonitrile): 3: 2, flow rate 1mL/min, detector DAD, detection wavelength 210nm, column temperature 30 ℃ C.) to determine the concentrations of R-2-phenoxypropionic acid and R-2- (4-hydroxyphenoxy) propionic acid. The results show that the concentration of R-2- (4-hydroxyphenoxy) propionic acid at the end of the reaction is 2.6 g/L; the mass conversion was 26%.
The composition of the slant medium, seed medium and fermentation medium in this example was the same as that in example 1.
Sequence listing
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<120> penicillium oxalicum ZJB16086 and application thereof in synthesis of R-2- (4-hydroxyphenoxy) propionic acid
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cgcgcccgcc gaagacacac aaacgaactc ttgtctgaag attgcagtct gagtacttga 180
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gcgaaatgcg ataagtaatg tgaattgcag aattcagtga atcatcgagt ctttgaacgc 300
acattgcgcc ccctggtatt ccggggggca tgcctgtccg agcgtcattg ctgccctcaa 360
gcacggcttg tgtgttgggc tctcgccccc cgcttccggg gggcgggccc gaaaggcagc 420
ggcggcaccg cgtccggtcc tcgagcgtat ggggcttcgt cacccgctct gtaggcccgg 480
ccggcgcccg ccggcgaaca ccatcaatct taaccaggtt gacctcggat caggtaggga 540
tacccgctga acttaagcat atcaata 567
Claims (7)
1. Penicillium oxalicum (Penicillium oxalicum) ZJB16086, deposited in China center for type culture Collection with the preservation number of CCTCC NO: M2018409, the preservation date of 2018, 6 months and 28 days, the preservation address: wuhan university, Wuhan, China, zip code 430072.
2. The use of the penicillium oxalicum ZJB16086 of claim 1 in the catalysis of R-2-phenoxypropionic acid to synthesize R-2- (4-hydroxyphenoxy) propionic acid.
3. The use according to claim 2, characterized in that said use is: taking fermentation liquor obtained by fermentation culture of penicillium oxalicum ZJB16086 as a catalyst and a reaction medium, adding a substrate R-2-phenoxypropionic acid, carrying out conversion reaction at the temperature of 25-30 ℃ and the table rotation speed of 150-.
4. The use according to claim 3, wherein the substrate has a final concentration of 10-50g/L in the fermentation broth, and the wet biomass content in the fermentation broth is 15-20 g/L.
5. The use according to claim 3, wherein the fermentation broth obtained by fermentative culture of Penicillium oxalicum ZJB16086 is prepared by the following method: inoculating penicillium oxalicum ZJB16086 to a fermentation culture medium, and performing fermentation culture at the temperature of 25-30 ℃ and the shaking table rotation speed of 150-; the fermentation medium comprises the following components: 5-20g/L glucose, 5-10g/L yeast extract, 5-10g/L ammonium sulfate, 0.5-1.0g/L magnesium sulfate heptahydrate, 0.05-0.12g/L manganese sulfate monohydrate, 1.5-2.5g/L potassium dihydrogen phosphate, 3.6-6.0g/L dipotassium hydrogen phosphate trihydrate, 1-5mL/L trace element liquid, deionized water as a solvent, and the pH value is 6.8; the trace element liquid comprises the following components: 2g/L of ferrous sulfate heptahydrate, 100mg/L of zinc sulfate tetrahydrate, 300mg/L of boric acid, 200mg/L of cobalt chloride hexahydrate, 10mg/L of copper chloride dihydrate, 20mg/L of nickel chloride hexahydrate and 30mg/L of sodium molybdate dihydrate, and deionized water is used as a solvent.
6. The use according to claim 5, characterized in that the fermentation medium consists of: 5g/L of glucose, 5g/L of yeast extract, 5g/L of ammonium sulfate, 0.5g/L of magnesium sulfate heptahydrate, 0.05g/L of manganese sulfate monohydrate, 1.5g/L of potassium dihydrogen phosphate, 3.6g/L of dipotassium hydrogen phosphate trihydrate, 1mL/L of trace element liquid, deionized water as a solvent, and 5M sodium hydroxide solution is used for adjusting the pH value to be 6.8.
7. The use according to claim 5, wherein the Penicillium oxalicum ZJB16086 is subjected to slant and seed culture before being inoculated into the fermentation medium: inoculating penicillium oxalicum ZJB16086 to a slant culture medium, and performing static culture at 28 deg.C for 6-7d to obtain slant strain; selecting thalli from the inclined plane, inoculating the thalli into a 250mL shaking flask filled with 50mL seed culture medium, and culturing at 28 ℃ and 150-; inoculating the seed solution into a 250mL shake flask containing 50mL fermentation medium in an inoculation amount with the volume concentration of 3% for fermentation culture; the slant culture medium comprises the following components: 200g/L of potato, 20g/L of glucose, 20g/L of agar and deionized water as a solvent, wherein the pH value is natural; the seed culture medium comprises the following components: potato 200g/L, glucose 20g/L, deionized water as solvent, and natural pH.
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