A method for preparing a chiral biphenyl-substituted propanol compound catalyzed by alcohol dehydrogenase
By using alcohol dehydrogenase catalysis to prepare chiral biphenyl substituted propanol compounds, the problems of complex synthesis methods and environmental pollution in existing technologies have been solved, enabling efficient and environmentally friendly industrial production.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- SUZHOU NOVARTIS PHARMA TECHONOLOGY CO LTD
- Filing Date
- 2024-11-22
- Publication Date
- 2026-05-29
AI Technical Summary
In the existing technology, the synthesis of chiral biphenyl substituted propanol compounds mainly relies on chemical synthesis methods, which have problems such as complex operation, high cost, and great environmental pollution.
Chiral biphenyl-substituted propanol compounds were prepared using alcohol dehydrogenase catalysis. The alcohol dehydrogenase was derived from Leifsonia sp. The engineered bacteria were constructed and induced to express the alcohol dehydrogenase. The catalytic reaction was carried out under specific reaction media and conditions, including the use of solubilizers, coenzymes, and cosubstrates.
A highly efficient and environmentally friendly method for the preparation of chiral biphenyl-substituted propanol compounds using alcohol dehydrogenase catalysis was achieved, with a conversion rate of up to 99% and an optical purity of over 99.5%, making the products suitable for industrial applications.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of bio-enzyme catalysis, and more specifically to a method for preparing high-optical-purity chiral biphenyl-substituted propanol compounds using alcohol dehydrogenases. Background Technology
[0002] Alcohol dehydrogenase (ADH), also known as ketone reductase (KRED) or carbonyl reductase (CRED), is considered a highly efficient and selective biocatalyst. It requires coenzymes such as NAD(H) or NADP(H) as hydrogen carriers and is widely found in animals, plants, and bacteria. Alcohol dehydrogenases selectively reduce carbonyl compounds to chiral alcohols, exhibiting advantages such as high substrate specificity, high selectivity, high conversion efficiency, mild reaction conditions, and low environmental pollution. Therefore, they have wide applications in the pharmaceutical and chemical industries.
[0003] In existing technologies, the synthesis of chiral biphenyl-substituted propanol compounds all employs chemical synthesis methods. It is necessary to develop enzymatic processes for the synthesis of chiral biphenyl-substituted propanol compounds. Summary of the Invention
[0004] To address the shortcomings of existing technologies, this invention provides a method for synthesizing chiral biphenyl-substituted propanol compounds using alcohol dehydrogenase catalysis. The biocatalytic preparation method provided by this invention is simple to operate, environmentally friendly, low-cost, and highly efficient, making it suitable for industrial applications.
[0005] This invention provides a method for preparing chiral biphenyl-substituted propanol compounds catalyzed by an alcohol dehydrogenase, wherein the alcohol dehydrogenase is derived from Leifsonia sp.
[0006] Furthermore, the amino acid sequence of the alcohol dehydrogenase is SEQ ID NO.1.
[0007] Amino acid sequence of alcohol dehydrogenase LsADH
[0008] SEQ ID NO.1
[0009] MAQYDVADRSAIVTGGGSGIGRAVALTLAASGAAVLVTDLNEEHAQAVVAEIEAAGGKAAALAGDVTDPAFGEASVAGANALAPLKIAVNNAGIGGEAATVGDYSLDSWRTVIEVNLNAVFYGMQ PQLKAMAANGGGAIVNMASILGSVGFANSSAYVTAKHALLGLTQNAALEYAADKVRVVAVGPGFIRTPLVEANLSADALAFLEGKHALGRLGEPEEVASLVAFLASDAASFITGSYHLVDGGYTAQ
[0010] Furthermore, the alcohol dehydrogenase is prepared using the following method:
[0011] (1) Synthesize the nucleotide sequence encoding the alcohol dehydrogenase described above and construct engineered bacteria;
[0012] (2) Induce culture of engineered bacteria, collect bacterial cells by centrifugation, and then prepare the required enzyme preparation;
[0013] The nucleotide sequence of the alcohol dehydrogenase is SEQ ID NO.2.
[0014] Nucleotide sequence of alcohol dehydrogenase LsADH
[0015] SEQ ID NO.2
[0016] atggctcagtacgacgtcgccgaccggtccgcgatcgtgaccggaggcggctcgggcatcgggcgcgccgtggcgctcactctcgcggcgagcg gcgcagccgtcctcgtcaccgacctgaacgaggagcacgcgcaggccgtcgtggccgagatcgaggccgcgggcggtaaggccgccgcgctcgcg ggcgacgtgaccgaccccgcgttcggcgaggcgagcgtcgccggggcgaacgctctcgcgcccctcaagatcgcggtcaacaacgcgggcatcg gcggcgaggccgccacggtcggcgactactcgctcgacagctggcgcacggtgatcgaggtcaacctcaacgccgtgttctacgggatgcagccg cagctgaaggccatggccgccaacggcggcggtgcgatcgtcaacatggcgtccatcctgggaagcgtcggcttcgccaactcgtcggcctacg tcacggccaagcacgcgctgctcggtctcacccagaacgccgcgctcgagtacgccgccgacaaggtgcgcgtcgtcgcggtcggccccggcttc atccgcaccccgctcgtggaggccaacctctccgccgacgcgctggcgttcctcgagggcaagcacgccctcggccgcctgggcgagccggaag aggtcgcctcgctggtcgcgttcctcgcctccgacgccgcgagcttcatcaccggcagctaccacctggtggacggcggctacaccgcccagtga
[0017] Furthermore, by adding a cosolvent, a substrate of biphenyl-substituted acetone, a co-substrate, a coenzyme, and an alcohol dehydrogenase to the reaction medium, chiral biphenyl-substituted propanol compounds can be obtained, as shown in the following reaction formula:
[0018]
[0019] Wherein, R can be hydrogen, halogen, alkyl, alkoxy, hydroxyl, etc. Preferably, R is chlorine, and the reaction formula is as follows:
[0020]
[0021] Furthermore, the reaction medium is triethanolamine-hydrochloric acid buffer, Tris-HCl buffer, phosphate buffer, borate-borax buffer, preferably Tris-HCl buffer.
[0022] Furthermore, the catalytic reaction temperature is 25-60℃, and more preferably, the reaction temperature is 55℃.
[0023] Furthermore, the initial reaction pH value is 6-10, and more preferably, the initial reaction pH value is 7.5.
[0024] Furthermore, the co-solvent is preferably dimethyl sulfoxide, and the amount added is 5-40% (V / V) of the reaction system, preferably 20% (V / V).
[0025] Furthermore, the co-substrate is isopropanol, and the preferred addition amount is isopropanol:substrate = 1:1 (V / W).
[0026] Furthermore, the coenzyme is NAD. + The addition amount is 0.5-5g / L, preferably 1g / L.
[0027] Furthermore, the enzyme-to-substrate ratio (the mass ratio of bacterial cells to substrate) is 1 / 5 to 1 / 150, preferably 1 / 20.
[0028] Furthermore, the concentration of the substrate is 10-200 g / L, preferably 100 g / L.
[0029] The alcohol dehydrogenase of this invention selectively reduces carbonyl compounds to chiral alcohols. The alcohol dehydrogenase from *Leifsonia* sp. catalyzes the formation of chiral biphenyl-substituted propanol compounds from biphenyl-substituted acetones. This alcohol dehydrogenase can catalyze substrate reactions at concentrations of 100 g / L with a conversion rate greater than 99%. The alcohol dehydrogenase of this invention exhibits strong catalytic activity towards ketone substrates and can react with high concentrations of substrates. The alcohol dehydrogenase of this invention has a strong affinity for its substrates, thus enabling large-scale industrial production of enzyme-catalyzed preparations.
[0030] Compared with the prior art, the present invention has the following beneficial effects:
[0031] (1) This invention first constructs a genetically engineered *E. coli* containing the *Leifsonia sp.* alcohol dehydrogenase gene, then cultivates and induces the expression of the alcohol dehydrogenase, and applies it to catalyze the synthesis of chiral biphenyl-substituted propanol compounds from biphenyl-substituted acetone. The alcohol dehydrogenase catalyzes a reaction with a feed concentration of 100 g / L, an enzyme-to-substrate ratio (bacterial mass to substrate mass ratio) of 1 / 20, a feed conversion rate >99%, and a product optical purity >99.5%.
[0032] (2) The alcohol dehydrogenase in this invention has significant catalytic activity, substrate affinity and regioselectivity for biphenyl-substituted acetones, and is highly tolerant to substrates and products. It has high feed concentration, low enzyme dosage, high yield, low waste and low cost, showing good industrialization potential. Detailed Implementation
[0033] To further understand the present invention, a method for preparing chiral biphenyl-substituted propanol compounds catalyzed by alcohol dehydrogenase, provided by the present invention, is described in detail below with reference to examples. It should be understood that these examples are described only to further illustrate the features of the present invention and are not intended to limit the scope of the invention or the scope of the claims.
[0034] Analytical methods, 1) Conversion rate determination by HPLC: Column: ZORBAX Eclipse XDB-C18 4.6mm×150mm×5μm, column temperature: 25℃, detection wavelength: UV238 nm. Mobile phase A: 30mM pH6.80 potassium dihydrogen phosphate-acetonitrile (90:10, V / V), mobile phase B: acetonitrile, flow rate: 1.0mL / min, gradient:
[0035] Table 1. HPLC mobile phase gradient
[0036] Time (min) Mobile phase A (%, V / V) Mobile phase B (%, V / V) 0 22 78 8 22 78 15 22 78 15.01 55 45 20 55 45 25 55 45
[0037] 2) Chiral HPLC analysis method: Column: CHIRALPAK.AD-H 5μm×4.6mmI.D.×250mmL, mobile phase: n-hexane:isopropanol (9:1, V / V), time: 20min, flow rate: 1mL / min, detection wavelength: UV 254nm.
[0038] Example 1: Preparation of alcohol dehydrogenase LsADH
[0039] 1. Construction of recombinant engineered bacteria producing alcohol dehydrogenase LsADH
[0040] Through enzyme information mining and screening, the alcohol dehydrogenase LsADH from Leifsonia sp. was obtained, and its amino acid sequence is shown in SEQ ID NO.1. The corresponding gene sequence was synthesized using the whole genome and is shown in SEQ ID NO.2. NdeI and EcoRI sites were introduced at both ends of the gene, and the plasmid was cloned into the pET21a vector. The constructed plasmid was chemically transformed into E. coli BL21(DE3) competent cells, plated on LB agar plates containing 50 μg / ml ampicillin resistance, and cultured at 37°C for 8–12 h. Single clones were picked to obtain a genetically engineered strain capable of inducibly expressing alcohol dehydrogenase.
[0041] 2. Induced expression of alcohol dehydrogenase LsADH
[0042] The strain expressing alcohol dehydrogenase was inoculated into LB medium containing Amp (0.1 mL of 50 g / L antibiotic aqueous solution in 100 mL of medium) and cultured overnight at 37°C and 200 rpm. Then, 1% (1 mL of seed culture in 100 mL of medium) was transferred into fermentation medium containing Amp (0.1 mL of 50 g / L antibiotic aqueous solution in 100 mL of medium) and cultured for another 3 h at 37°C and 200 rpm. Finally, 0.05 mM IPTG (0.025 mL of 50 g / L IPTG aqueous solution in 100 mL of medium) was added, and induction was performed at 25°C and 200 rpm for 20 h. The fermentation medium composition was: 3 g / L glycerol, 15 g / L peptone, 0.5 g / L NaH2PO4, 10 mM FeSO4, pH 7.0.
[0043] 3. Preparation of LsADH enzyme solution
[0044] The alcohol dehydrogenase cells obtained after centrifuging the fermentation broth were resuspended evenly in 0.1M pH7.0 Tris-HCl buffer to a cell concentration of 200 g / L. The cells were then disrupted in an ultrasonic disruptor (400W, 2s operation, 6s rest, 30min sonication) to obtain the cell disruption solution, which is the enzyme solution.
[0045] Example 2: Reaction test for the synthesis of chiral biphenyl-substituted propanol compounds catalyzed by alcohol dehydrogenase.
[0046] A 1 mL reaction system contains 5 g / L biphenyl-substituted acetone, 10% (v / v) dimethyl sulfoxide, and 1 g / L NAD. + 20 μL of the alcohol dehydrogenase LsADH enzyme solution prepared in Example 1 (cell volume 0.004 g), 2% (V / V) isopropanol, was added, and the volume was adjusted to 1 mL with 0.1 M, pH 8.0 Tris-HCl. The reaction was carried out at 30 °C and 1000 rpm for 24 h. Samples were taken for analysis, and the results are shown in Table 2.
[0047] Table 2 Results of the synthesis of chiral biphenyl-substituted propanol compounds catalyzed by alcohol dehydrogenase.
[0048]
[0049]
[0050] Example 3: Synthesis of (S)-1-(4-biphenyl)-3-chloro-2-propanol catalyzed by alcohol dehydrogenase in different reaction media
[0051] 10 mL reaction system, 20 g / L starting material, 10% (v / v) dimethyl sulfoxide, 1 g / L NAD + 20 μL of the alcohol dehydrogenase LsADH enzyme solution prepared in Example 1 (0.004 g of bacterial cells), 2% (v / v) isopropanol, were added, and the volume was adjusted to 10 mL using different 0.1 M, pH 8.0 buffer solutions. The reaction was carried out at 30 °C for 1 h. As shown in Table 3, the highest conversion rate was achieved when Tris-HCl buffer was used as the reaction medium.
[0052] Table 3 Results of alcohol dehydrogenase-catalyzed reactions in different reaction media over 1 hour
[0053] reaction medium Conversion rate Product ee% value Triethanolamine-hydrochloric acid buffer 7.5％ 99.7％ Tris-HCl buffer 8.1％ 99.8％ Phosphate buffer 7.3％ 99.8％ Boric acid-borax buffer solution 7.1％ 99.7％
[0054] Example 4: Synthesis of (S)-1-(4-biphenyl)-3-chloro-2-propanol catalyzed by alcohol dehydrogenase at different temperatures
[0055] 10 mL reaction system, 20 g / L starting material, 10% (v / v) dimethyl sulfoxide, 1 g / L NAD + 20 μL of the alcohol dehydrogenase LsADH enzyme solution prepared in Example 1 (cell volume 0.004 g), 2% (v / v) isopropanol, and the volume was adjusted to 10 mL with 0.1 M, pH 8.0 Tris-HCl buffer. The reaction was carried out at different temperatures of 25-60 °C for 1 h. As shown in Table 4, the conversion rate was the highest at a reaction temperature of 55 °C.
[0056] Table 4 Results of alcohol dehydrogenase-catalyzed reaction at different temperatures for 1 h
[0057]
[0058] Example 5: Synthesis of (S)-1-(4-biphenyl)-3-chloro-2-propanol catalyzed by alcohol dehydrogenase under different initial pH conditions
[0059] 10 mL reaction system, 20 g / L starting material, 10% (v / v) dimethyl sulfoxide, 1 g / L NAD + 20 μL of the alcohol dehydrogenase LsADH enzyme solution prepared in Example 1 (0.004 g of bacterial cells) was added, and 2% (v / v) isopropanol was added. The solution was then diluted to 10 mL with 0.1 M Tris-HCl buffer at pH 6-10 and reacted at 55 °C for 1 h. As shown in Table 5, the highest conversion rate was achieved at a reaction pH of 7.5.
[0060] Table 5 Results of alcohol dehydrogenase-catalyzed reactions at different initial pH levels over 1 hour
[0061] pH Conversion rate Product ee% value 6 1.2％ 99.6％ 6.5 2.3％ 99.8％ 7 11.5％ 99.5％ 7.5 19.5％ 99.8％ 8 14.6％ 99.8％ 8.5 10.8％ 99.7％ 9 5.5％ 99.6％ 9.5 3.3％ 99.8％ 10 3.3％ 99.8％
[0062] Example 6 Synthesis of (S)-1-(4-biphenyl)-3-chloro-2-propanol catalyzed by alcohol dehydrogenase under different cosolvents
[0063] 10 mL reaction system, 20 g / L starting material, 10% (v / v) different co-solvents, 1 g / L NAD + 20 μL of the alcohol dehydrogenase LsADH enzyme solution prepared in Example 1 (cell volume 0.004 g) was added to 2% (v / v) isopropanol and diluted to 10 mL with 0.1 M, pH 7.5 Tris-HCl buffer. The reaction was carried out at 55 °C for 1 h. As shown in Table 6, the highest conversion rate was achieved when dimethyl sulfoxide was used as the co-solvent.
[0064] Table 6 Results of alcohol dehydrogenase-catalyzed reaction under different cosolvents for 1 h
[0065] Types of cosolvents Conversion rate Product ee% value No cosolvent added 18.1％ 99.8％ Dimethyl sulfoxide 19.6％ 99.7％ N,N-Dimethylformamide 3.5％ 99％ methanol 2.5％ 99％ ethanol 1.3％ 99.8％ Acetonitrile 1.9％ 99.3％ Isopropanol 18.8％ 99.2％ Tetrahydrofuran 1.6％ 99.4％ 1,4-Dioxane 0％ / 2-Methyltetrahydrofuran 2.5％ 99.5％
[0066] Example 7: Synthesis of (S)-1-(4-biphenyl)-3-chloro-2-propanol catalyzed by alcohol dehydrogenase under different amounts of cosolvent.
[0067] 10 mL reaction system, 20 g / L raw material, different amounts of dimethyl sulfoxide, 1 g / L NAD + 20 μL of the alcohol dehydrogenase LsADH enzyme solution prepared in Example 1 (cell volume 0.004 g), 2% (v / v) isopropanol, and the volume was adjusted to 10 mL with 0.1 M, pH 7.5 Tris-HCl buffer. The reaction was carried out at 55 °C for 1 h. As shown in Table 7, the highest conversion efficiency was achieved when the co-solvent dimethyl sulfoxide was added at 20%.
[0068] Table 7 Results of alcohol dehydrogenase catalytic reaction after 1 h under different cosolvent addition amounts
[0069] Amount of cosolvent added (V / V) Conversion rate Product ee% value 5％ 14.8％ 99.8％ 10％ 19.3％ 99.7％ 20％ 20.9％ 99.8％ 30％ 18.6％ 99.5％ 40％ 14.8％ 99.5％
[0070] Example 8: Synthesis of (S)-1-(4-biphenyl)-3-chloro-2-propanol catalyzed by alcohol dehydrogenase under different amounts of cosubstrate.
[0071] 10 mL reaction system, 20 g / L starting material, 20% (v / v) dimethyl sulfoxide, 1 g / L NAD + 20 μL of the alcohol dehydrogenase LsADH enzyme solution prepared in Example 1 (0.004 g of bacterial cells) was added, with different amounts of the co-substrate isopropanol added (1-10% (V / V). The solution was then diluted to 10 mL with 0.1 M, pH 7.5 Tris-HCl buffer and reacted at 55 °C for 1 h. As shown in Table 8, the catalytic efficiency was highest when the isopropanol addition was 2% (V / V), i.e., isopropanol:substrate = 1:1 (V / W).
[0072] Table 8 Results of alcohol dehydrogenase-catalyzed reaction for 1 h under different amounts of isopropanol cosubstrate.
[0073] Isopropanol addition (V / V) Conversion rate Product ee% value 1％ 14.7％ 99.8％ 2％ 20.2％ 99.5％ 5％ 19.9％ 99.3％ 7％ 19.6％ 99.8％ 10％ 19％ 99.6％
[0074] Example 9: Synthesis of (S)-1-(4-biphenyl)-3-chloro-2-propanol catalyzed by alcohol dehydrogenase under different coenzyme addition amounts
[0075] A 10 mL reaction system, 20 g / L raw material, 20% (v / v) dimethyl sulfoxide, and different amounts (0.5-5 g / L) of coenzyme NAD+. + 20 μL of the alcohol dehydrogenase LsADH enzyme solution prepared in Example 1 (cell volume 0.004 g), 2% (v / v) isopropanol, and the volume was adjusted to 10 mL with 0.1 M, pH 7.5 Tris-HCl buffer. The reaction was carried out at 55 °C for 1 h. As shown in Table 9, NAD... + When the addition amount is 1-5g / L, the increase in conversion rate is not significant. Considering the overall cost, the coenzyme dosage is determined to be 1g / L.
[0076] Table 9 Different coenzymes NAD + Results of alcohol dehydrogenase-catalyzed reaction for 1 hour at the specified addition amount
[0077] <![CDATA[NAD + (g / L)]]> Conversion rate Product ee% value 0.5 13％ 99.8％ 1 20.9％ 99.1％ 2.5 21.2％ 99.5％ 3.5 21.4％ 99.3％ 5 21.8％ 99.5％
[0078] Example 10 Synthesis of (S)-1-(4-biphenyl)-3-chloro-2-propanol catalyzed by alcohol dehydrogenase under different enzyme ratios
[0079] 10 mL reaction system, 2-60 g / L starting material, 20% (v / v) dimethyl sulfoxide, 0.2-6% (v / v) isopropanol, 1 g / L coenzyme NAD. + 20 μL of the alcohol dehydrogenase LsADH enzyme solution prepared in Example 1 (cell mass 0.004 g) was brought to a final volume of 10 mL with 0.1 M, pH 7.5 Tris-HCl buffer and reacted at 55 °C for 1 h. As shown in Table 10, considering both enzyme usage cost and conversion rate, the enzyme-to-substrate ratio (mass ratio of cell mass to substrate) was determined to be 1 / 20.
[0080] Table 10 Results of alcohol dehydrogenase-catalyzed reactions at different enzyme ratios over 1 hour
[0081]
[0082]
[0083] Example 11 Synthesis of (S)-1-(4-biphenyl)-3-chloro-2-propanol catalyzed by alcohol dehydrogenase under different substrate concentrations
[0084] A 10 mL reaction system was used, with varying amounts of raw materials, an enzyme-to-substrate ratio (bacterial cell mass to substrate mass ratio) of 1 / 20, and 20% (v / v) dimethyl sulfoxide and coenzyme NAD+. +1 g / L, different amounts of the alcohol dehydrogenase LsADH enzyme solution prepared in Example 1, different amounts of isopropanol, were diluted to 10 mL with Tris-HCl buffer at pH 7.5 and reacted at 55 °C for 24 h. Table 11 shows that when the feed concentration is 100 g / L, 250 μL of enzyme solution (0.05 g of bacterial cells), and the coenzyme NAD... + 1 g / L, 10% isopropanol (V / V), conversion rate 99.3%, product 99.6% ee.
[0085] Table 11 Results of alcohol dehydrogenase-catalyzed reaction at different substrate concentrations after 24 h
[0086]
[0087] The above are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.
Claims
1. A method for preparing chiral biphenyl-substituted propanol compounds catalyzed by alcohol dehydrogenase, characterized in that: The reaction of biphenyl-substituted acetones to generate chiral biphenyl-substituted propanols is catalyzed by an alcohol dehydrogenase derived from Leifsonia sp., and the reaction formula is shown below: Wherein, R is hydrogen, halogen, alkyl, alkoxy, or hydroxyl.
2. The preparation method according to claim 1, characterized in that, R is chlorine, and the product prepared is (S)-1-(4-biphenyl)-3-chloro-2-propanol.
3. The method for preparing chiral biphenyl-substituted propanol compounds catalyzed by alcohol dehydrogenase according to claim 1, characterized in that, The amino acid sequence of the alcohol dehydrogenase is SEQ ID NO.
1.
4. The method for preparing chiral biphenyl-substituted propanol compounds catalyzed by alcohol dehydrogenase according to claim 1 or 2, characterized in that, This includes the steps for preparing alcohol dehydrogenase: (1) Synthesize the nucleotide sequence encoding the alcohol dehydrogenase and construct engineered bacteria; (2) Induce culture of engineered bacteria, collect bacterial cells by centrifugation, and then prepare the required enzyme preparation; The nucleotide sequence is SEQ ID NO.
2.
5. The method for preparing chiral biphenyl-substituted propanol compounds catalyzed by alcohol dehydrogenase according to claim 1 or 2, characterized in that, In the reaction medium, a cosolvent, a co-substrate, a coenzyme, and a biphenyl-substituted acetone are added, followed by the addition of an alcohol dehydrogenase to carry out the reaction, yielding a chiral biphenyl-substituted propanol compound.
6. The method for preparing chiral biphenyl-substituted propanol compounds catalyzed by alcohol dehydrogenase according to claim 5, characterized in that, The reaction media are triethanolamine-hydrochloric acid buffer, Tris-HCl buffer, phosphate buffer, and borate-borax buffer.
7. The method for preparing chiral biphenyl-substituted propanol compounds catalyzed by alcohol dehydrogenase according to claim 5, characterized in that, The reaction temperature is 25-60℃, and the initial reaction pH is 6-10.
8. The method for preparing chiral biphenyl-substituted propanol compounds catalyzed by alcohol dehydrogenase according to claim 5, characterized in that, The cosolvent is dimethyl sulfoxide, added at 5-40% (v / v) of the reaction system; the cosubstrate is isopropanol, with an isopropanol:substrate ratio of 1:1 (v / w); the coenzyme is NAD+. + The addition amount is 0.5-5g / L.
9. The method for preparing chiral biphenyl-substituted propanol compounds catalyzed by alcohol dehydrogenase according to claim 5, characterized in that, The enzyme-to-substrate ratio (the ratio of cell mass to substrate mass) is 1 / 5 to 1 / 150.
10. The method for preparing chiral biphenyl-substituted propanol compounds catalyzed by alcohol dehydrogenase according to claim 5, characterized in that, The concentration of the substrate, biphenyl-substituted acetone, is 10-200 g / L.