A process for the in situ extractive fermentation production of furfurylamine

Abstract

The application discloses a method for preparing furfurylamine by in-situ extraction fermentation, and belongs to the technical field of bio-chemical industry. In order to solve the problems of strong raw material and product inhibition effect, high concentration of by-products and low concentration of product in the preparation of furfurylamine by biological transamination catalysis, the application provides a method for preparing furfurylamine by in-situ extraction fermentation, specifically, fermentation culture and induction are carried out on a strain, then furfural, an amino donor and glucose are added in a flow manner to carry out a transamination reaction, and an external flow addition of an extractant is carried out to carry out extraction fermentation. The application firstly reduces the generation of by-products by using the biological fermentation technology, simultaneously couples the extraction separation technology to separate furfurylamine in-situ, effectively solves the problems of biological toxicity and multiple side reactions faced by the biological transamination, and realizes the biological fermentation of high-concentration furfurylamine.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of bio-chemical industry, and relates to a method for preparing furfurylamine by in-situ extraction fermentation, in particular to a green and efficient method for preparing furfurylamine by in-situ extraction and intensified bio-fermentation. BACKGROUND

[0002] Furfurylamine is a primary amine compound derived from furfural, which has various applications in modern chemistry and industry. Due to the advantages of high selectivity, high catalytic efficiency and mild reaction conditions, biological catalysis has become the research frontier of the preparation of bio-based furfurylamine. For example, in 2017, Alice et al. (Dunbabin A, Subrizi F, Ward J M, et al. Furfurylamines from biomass: transaminase catalysed upgrading of furfurals[J]. Green Chem, 2017, 19: 397-404) used transaminase (CV-TAMs) derived from Chromobacteium violaceum (DSM30191) to catalyze furfural to prepare furfurylamine. In 2021, Li Qing et al. (Li Q, Di J, Liao X, et al. Exploration of benign deep eutectic solvent-water systems for the highly efficient production of furfurylamine from sugarcane bagasse via chemoenzymatic cascade catalysis[J]. Green Chem, 2021, 23: 8154-8168) used transaminase (CV-TAMs) and alanine dehydrogenase (AlaDH) in a two-phase system of choline chloride / ethylene glycol-water, and used ammonium chloride as an amino donor to catalyze the synthesis of furfurylamine. At present, the preparation of furfurylamine by transaminase catalysis of furfural is still at the laboratory research level, and it has the problems of strong inhibition effect of raw materials and products, high concentration of by-products and low concentration of products, which restricts the biological preparation of high-concentration furfurylamine, and the accumulation concentration of furfurylamine is always maintained below 10 g / L.

[0003] Reaction and separation coupling process is developed to solve the problem of low product concentration and low conversion rate caused by product inhibition, and in-situ extraction fermentation is a typical reaction and separation coupling process, which can reduce product inhibition by instant separation of product during fermentation, and for fermentation process with by-products, extraction fermentation can also improve the ratio of product and by-product, thereby further improving the product yield. At present, the biological preparation of furfurylamine is still in the laboratory flask stage, and there is a lack of effective in-situ extraction fermentation technology of furfurylamine.

[0004] On this basis, in view of the problems of strong raw material and product inhibition effect, high by-product concentration and low product concentration existing in the preparation of furfurylamine by biological transamination catalysis, the generation of by-products is reduced by optimizing the biological fermentation technology, and a new type of in-situ extraction fermentation technology of furfurylamine is developed by coupling extraction separation technology, which is expected to solve the problems of biological toxicity and multiple side reactions, and realize the biological fermentation of high-concentration furfurylamine. SUMMARY

[0005] In order to overcome the problems of strong raw material and product inhibition effect and low product concentration existing in the present biological transamination, the application provides a method for preparing furfurylamine by in-situ extraction fermentation, which realizes in-situ separation of furfurylamine by using in-situ extraction fermentation technology for the first time, eliminates product inhibition, and realizes high-concentration furfurylamine fermentation preparation.

[0006] The method for preparing furfurylamine by in-situ extraction fermentation provided by the application is carried out according to the following steps:

[0007] Step 1: inoculating activated transaminase-producing recombinant engineering strain into fermentation medium containing glucose for fermentation;

[0008] Step 2: after adding an inducer to induce fermentation, adding furfural, amino donor and glucose for transamination reaction, and then adding an extractant exogenously for extraction fermentation, wherein the extractant can extract furfurylamine produced in the fermentation broth in-situ;

[0009] Step 3: after the fermentation is completed, recovering the extractant, and distilling under reduced pressure to obtain furfurylamine.

[0010] In an embodiment of the application, the transaminase-producing recombinant engineering strain in step 1 is E.coli LYJ201.

[0011] In an embodiment of the application, the inoculation amount of the recombinant engineering strain in step 1 is 1% to 10% of the volume of the fermentation medium.

[0012] In an embodiment of the application, the glucose concentration in the fermentation medium in step 1 is 10 to 20 g / L.

[0013] In an embodiment of the present application, the components of the fermentation medium in step 1, except glucose, are 5-10 g / L K2HPO4·3H2O, 1-3 g / L citric acid monohydrate, 0.1-0.5 g / L ferric ammonium citrate, 2-3 g / L ammonium sulfate, and 0.1-0.3 g / L MgSO4·7H2O.

[0014] In an embodiment of the present application, the fermentation culture conditions in step 1 are 25-40℃, 100-1000 rpm, and 10-15 h.

[0015] In an embodiment of the present application, the induction fermentation in step 2 refers to adding an inducer IPTG to a final concentration of 50-200 μM when OD 600 is 10-20, and inducing at 25-35℃, 100-1000 rpm for 2-6 h.

[0016] In an embodiment of the present application, the feeding speed of the furfural in step 2 is 0.5-5 g / (L·h), the feeding speed of the amino donor is 0.5-6 g / (L·h), and the feeding speed of the glucose is 1.5-15 g / (L·h).

[0017] In an embodiment of the present application, the amino donor in step 2 includes any one of ammonium chloride and ammonium sulfate.

[0018] In an embodiment of the present application, the extractant in step 2 includes any one of trialkyl phosphine oxide, tricaprylyl phosphate, tributyl phosphate, n-octane, and dodecane.

[0019] In an embodiment of the present application, the extractant is added at a volume fraction of 20%-50% based on the volume of the fermentation broth, and the feeding speed is 0.03-0.18 V / h.

[0020] In an embodiment of the present application, the extraction fermentation culture conditions in step 2 are 25-40℃, 100-1000 rpm, and 10-15 h.

[0021] In an embodiment of the present application, the extractant is recovered by any one of centrifugation and liquid separation.

[0022] The present application has the following beneficial effects:

[0023] The method of the present application aims at the problems of strong inhibition effect of raw materials and products, high concentration of by-products and low concentration of products existing in the preparation of furfurylamine by biological transamination catalysis, reduces the generation of by-products by using biological fermentation technology, simultaneously in-situ separates furfurylamine by coupling extraction separation technology, effectively solves the problems of biological toxicity and multiple side reactions faced by biological transamination, and realizes the biological fermentation of high-concentration furfurylamine. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 The H NMR results of the furfurylamine obtained in Example 5 are as follows: 1 H NMR results;

[0025] Figure 2 The H NMR results of the furfurylamine obtained in Example 5 are as follows: 13 C NMR results. DETAILED DESCRIPTION

[0026] In order to make the purpose, technical scheme and advantages of the present application more clear, the present application will be further described in detail below in combination with specific implementation modes and the drawings of the specification. The experimental methods used in the following examples are all conventional methods unless otherwise specified. The materials, reagents, methods and instruments used are all conventional materials, reagents, methods and instruments in the art unless otherwise specified, and can be obtained by commercial channels by those skilled in the art.

[0027] The recombinant strain used in the following examples and comparative examples is E. coli LYJ201, which is disclosed in the invention patent with application number 202211645773.6. The recombinant strain is obtained by knocking out the DNA binding transcriptional dual regulator arcA and overexpressing the transaminase gene OATA, the alanine dehydrogenase gene ALD and the T7 promoter connected carbon storage regulator CsrB gene in the E. coli BL21 (DE3) strain. The transaminase gene OATA is derived from the human Ochrobactrum anthropi, and the gene sequence is shown in SEQ ID NO. 3. The alanine dehydrogenase gene ALD is derived from Bacillus subtilis, and the Genebank ID is 936557. The sequence of the csrB gene is shown in SEQ ID NO. 1. The sequence of the DNA binding transcriptional dual regulator arcA is shown in SEQ ID NO. 4. The nucleotide sequence of the T7 promoter is shown in SEQ ID NO. 2.

[0028] The culture medium involved in the following examples and comparative examples is as follows:

[0029] LB medium: Typtone 10 g / L, Yeast Extract 5 g / L, NaCl 10 g / L.

[0030] M9 medium: NH4CI 1.0 g / L, Na2HPO4·12H2O 15.2 g / L, KH2PO4 3.0 g / L, NaCI 0.5 g / L.

[0031] Comparative Example 1:

[0032] This comparative example utilizes recombinant strain E. coli LYJ201 to prepare furfurylamine by transamination fermentation, which is specifically carried out according to the following steps:

[0033] Step 1: inoculate the recombinant strain single colony on the solid LB plate in the LB liquid medium, add 1 ‰ chloramphenicol, and culture at 37℃, 200 rpm for 9 h to obtain the primary seed liquid;

[0034] Step 2: inoculate the primary seed liquid into 250 mL fermentation shake flasks containing 75 mL M9 medium at a 2% (wt) inoculation amount, add 1 ‰ chloramphenicol when inoculating the primary seed liquid, and culture at 180 rpm, 37℃ for 8 h to obtain the secondary seed liquid;

[0035] Step 3: inoculate the secondary seed liquid into a 5 L fermentation tank containing 1.5 L M9 medium at a 5% (wt) inoculation amount for fermentation, add 1 ‰ chloramphenicol when inoculating the seed liquid, and culture at 500 rpm, 37℃ for 11 h;

[0036] Step 4: when the fermentation culture reaches OD 600 15, then add the inducer IPTG to a final concentration of 50 μM, induce expression at 30℃, 500 rpm for 3 h, then flow add the substrate furfural at a flow rate of 2 g / (L.h), flow add the amino donor (ammonium chloride) at a flow rate of 3 g / (L.h), and flow add glucose at a flow rate of 6 g / (L.h), and transamination reaction for 9 h to prepare furfurylamine. Centrifuge the sample, dilute the supernatant 50 times, and then detect the furfurylamine concentration by HPLC. The furfurylamine concentration is 12.1 g / L.

[0037] Comparative Example 2:

[0038] This comparative example utilizes recombinant strain E. coli LYJ201 to prepare furfurylamine by transamination fermentation, which is specifically carried out according to the following steps:

[0039] Step 1: inoculate the recombinant strain single colony on the solid LB plate in the LB liquid medium, add 1 ‰ chloramphenicol, and culture at 37℃, 200 rpm for 9 h to obtain the primary seed liquid;

[0040] Step 2: The first-stage seed liquid was inoculated into 250 mL fermentation flasks containing 75 mL M9 medium at a 2% (wt) inoculation amount, and 1‰ chloramphenicol was added when the first-stage seed liquid was inoculated, 180 rpm, 37°C for 8 h, to obtain a second-stage seed liquid;

[0041] Step 3: The second-stage seed liquid was inoculated into a 5 L fermentation tank containing 1.5 L M9 medium at a 5% (wt) inoculation amount for fermentation, and 1‰ chloramphenicol was added when the seed liquid was inoculated, 500 rpm, 37°C for 11 h;

[0042] Step 4: The fermentation was cultured to OD 600 was 15, and then an inducer IPTG was added to a final concentration of 50 μM, 30°C, 500 rpm, and expression was induced for 3 h, and then the substrate furfural was fed at a flow rate of 2 g / (L.h), the amino donor (ammonium chloride) was fed at a flow rate of 6 g / (L.h), and glucose was fed at a flow rate of 6 g / (L.h), and transamination reaction was performed for 9 h to prepare furfurylamine. After centrifugation, the supernatant was diluted 50 times, and HPLC detection was performed, and the furfurylamine concentration was 15.1 g / L.

[0043] Example 1:

[0044] In this example, the extractant tributyl phosphate was used to strengthen the transamination fermentation of the recombinant strain E. coli LYJ201 to prepare furfurylamine, and the following steps were performed:

[0045] Step 1: A single colony of the recombinant strain on a solid LB plate was inoculated in LB liquid medium, and 1‰ chloramphenicol was added, 37°C, 200 rpm for 8 h, to obtain a first-stage seed liquid;

[0046] Step 2: The first-stage seed liquid was inoculated into 250 mL fermentation flasks containing 75 mL M9 medium at a 2% (wt) inoculation amount, and 1‰ chloramphenicol was added when the first-stage seed liquid was inoculated, 180 rpm, 37°C for 9 h, to obtain a second-stage seed liquid;

[0047] Step 3: The second-stage seed liquid was inoculated into a 5 L fermentation tank containing 1.5 L M9 medium at a 5% (wt) inoculation amount for fermentation, and 1‰ chloramphenicol was added when the seed liquid was inoculated, 500 rpm, 37°C for 10 h;

[0048] Step 4: The fermentation was cultured to OD 600The recombinant strain E. coli LYJ201 was inoculated with a single colony on a solid LB plate in LB liquid medium with 1‰ chloramphenicol, and cultured at 37°C and 200 rpm for 8 h to obtain a primary seed solution. The primary seed solution was inoculated into 250 mL fermentation flasks containing 75 mL M9 medium at a 2% (wt) inoculation amount, and 1‰ chloramphenicol was added. The flasks were cultured at 180 rpm and 37°C for 9 h to obtain a secondary seed solution. The secondary seed solution was inoculated into a 5 L fermentation tank containing 1.5 L M9 medium at a 5% (wt) inoculation amount, and 1‰ chloramphenicol was added. The tank was cultured at 500 rpm and 37°C for 10 h. When the optical density (OD) of the culture reached 16, an inducer IPTG was added to a final concentration of 50 μM, and the culture was induced at 30°C and 500 rpm for 5 h. Then, the substrate furfural was fed at a flow rate of 4 g / (L.h), the amino donor (ammonium chloride) was fed at a flow rate of 6 g / (L.h), and glucose was fed at a flow rate of 12 g / (L.h). After the transamination reaction was performed for 4 h, the extractant trioctyl phosphate was fed at a flow rate of 150 mL / h for 3 h, and the transamination reaction was continued for 11 h to prepare furfurylamine. The sample was centrifuged, and the supernatant was diluted 50 times. The furfurylamine concentration was determined by HPLC to be 22.4 g / L.

[0049] Example 2

[0050] In this example, the extractant trioctyl phosphate was used to strengthen the transamination fermentation of the recombinant strain E. coli LYJ201 to prepare furfurylamine. The following steps were taken:

[0051] Step 1: The recombinant strain was inoculated with a single colony on a solid LB plate in LB liquid medium with 1‰ chloramphenicol, and cultured at 37°C and 200 rpm for 8 h to obtain a primary seed solution.

[0052] Step 2: The primary seed solution was inoculated into 250 mL fermentation flasks containing 75 mL M9 medium at a 2% (wt) inoculation amount, and 1‰ chloramphenicol was added. The flasks were cultured at 180 rpm and 37°C for 9 h to obtain a secondary seed solution.

[0053] Step 3: The secondary seed solution was inoculated into a 5 L fermentation tank containing 1.5 L M9 medium at a 5% (wt) inoculation amount, and 1‰ chloramphenicol was added. The tank was cultured at 500 rpm and 37°C for 10 h.

[0054] Step 4: When the optical density (OD) of the culture reached 16, an inducer IPTG was added to a final concentration of 50 μM, and the culture was induced at 30°C and 500 rpm for 5 h. Then, the substrate furfural was fed at a flow rate of 4 g / (L.h), the amino donor (ammonium chloride) was fed at a flow rate of 6 g / (L.h), and glucose was fed at a flow rate of 12 g / (L.h). After the transamination reaction was performed for 4 h, the extractant trioctyl phosphate was fed at a flow rate of 150 mL / h for 3 h, and the transamination reaction was continued for 11 h to prepare furfurylamine. The sample was centrifuged, and the supernatant was diluted 50 times. The furfurylamine concentration was determined by HPLC to be 22.4 g / L. 600

[0055] Example 3

[0056] In this example, the extractant trioctyl phosphate was used to strengthen the transamination fermentation of the recombinant strain E. coli LYJ201 to prepare furfurylamine. The following steps were taken: ​

[0057] Step 1: inoculate the single colony of the recombinant strain on the solid LB plate into LB liquid medium with 1‰ chloramphenicol, cultivate at 37℃, 200 rpm for 8 h, and obtain the primary seed liquid;

[0058] Step 2: inoculate the primary seed liquid into 250 mL fermentation shake flasks containing 75 mL M9 medium with 2% (wt) inoculation amount, supplement 1‰ chloramphenicol when inoculating the primary seed liquid, cultivate at 180 rpm, 37℃ for 9 h, and obtain the secondary seed liquid;

[0059] Step 3: inoculate the secondary seed liquid into a 5 L fermentation tank containing 1.5 L M9 medium with 5% (wt) inoculation amount for fermentation, supplement 1‰ chloramphenicol when inoculating the seed liquid, cultivate at 500 rpm, 37℃ for 10 h;

[0060] Step 4: cultivate to OD 600 is 16, then add the inducer IPTG to a final concentration of 50 μM, cultivate at 30℃, 500 rpm for 5 h to induce expression, then add the substrate furfural at a flow rate of 4 g / (L.h), add the amino donor (ammonium chloride) at a flow rate of 6 g / (L.h), add glucose at a flow rate of 12 g / (L.h), add the extractant n-octane at a flow rate of 150 mL / h for 3 h after the transamination reaction for 4 h, and continue the transamination reaction for 11 h to prepare furfurylamine. Centrifuge the sample, dilute the supernatant 50 times, and then detect the furfurylamine concentration by HPLC. The furfurylamine concentration is 22.3 g / L.

[0061] Example 4:

[0062] Example 4:

[0063] Step 1: inoculate the single colony of the recombinant strain on the solid LB plate into LB liquid medium with 1‰ chloramphenicol, cultivate at 37℃, 200 rpm for 8 h, and obtain the primary seed liquid;

[0064] Step 2: inoculate the primary seed liquid into 250 mL fermentation shake flasks containing 75 mL M9 medium with 2% (wt) inoculation amount, supplement 1‰ chloramphenicol when inoculating the primary seed liquid, cultivate at 180 rpm, 37℃ for 9 h, and obtain the secondary seed liquid;

[0065] Step 3: inoculate the secondary seed liquid into a 5 L fermentation tank containing 1.5 L M9 medium with 5% (wt) inoculation amount for fermentation, supplement 1‰ chloramphenicol when inoculating the seed liquid, cultivate at 500 rpm, 37℃ for 10 h;

[0066] Step 4: culture to OD 600 was 16, then inducer IPTG was added to a final concentration of 50 μΜ, 30°C, 500 rpm, expression was induced for 5 h, then substrate furfural was fed at a flow rate of 4 g / (L.h), amino donor (ammonium chloride) was fed at a flow rate of 5 g / (L.h), glucose was fed at a flow rate of 10 g / (L.h), after 4 h of transamination, extractant trialkylphosphine oxide was fed at a flow rate of 75 mL / h for 4 h, and the transamination reaction was continued for 12 h to prepare furfurylamine. The sample was centrifuged, the supernatant was diluted 50 times, and HPLC detection was performed, and the furfurylamine concentration was 27.2 g / L.

[0067] Example 5:

[0068] This example uses extractant trialkylphosphine oxide to strengthen the transamination fermentation of recombinant strain E. coli LYJ201 to prepare furfurylamine, which is specifically carried out according to the following steps:

[0069] Step 1: inoculate the recombinant strain single colony on a solid LB plate into LB liquid medium, add 1 ‰ chloramphenicol, and culture at 37°C, 200 rpm for 8 h to obtain a primary seed liquid;

[0070] Step 2: inoculate the primary seed liquid into 250 mL fermentation flasks containing 75 mL M9 medium at a 2% (wt) inoculation amount, add 1 ‰ chloramphenicol when inoculating the primary seed liquid, and culture at 180 rpm, 37°C for 8 h to obtain a secondary seed liquid;

[0071] Step 3: inoculate the secondary seed liquid into a 5 L fermentation tank containing 1.5 L M9 medium at a 5% (wt) inoculation amount for fermentation, add 1 ‰ chloramphenicol when inoculating the seed liquid, and culture at 500 rpm, 37°C for 11 h;

[0072] Step 4: culture to OD 600 was 16, then inducer IPTG was added to a final concentration of 50 μΜ, 30°C, 500 rpm, expression was induced for 5 h, then substrate furfural was fed at a flow rate of 4 g / (L.h), amino donor (ammonium chloride) was fed at a flow rate of 5 g / (L.h), glucose was fed at a flow rate of 10 g / (L.h), after 4 h of transamination, extractant trialkylphosphine oxide was fed at a flow rate of 75 mL / h for 4 h, and the transamination reaction was continued for 12 h to prepare furfurylamine. The sample was centrifuged, the supernatant was diluted 50 times, and HPLC detection was performed, and the furfurylamine concentration was 27.2 g / L. 1 H NMR results are shown in Figure 1 , 13 C NMR results are shown in Figure 2 .

[0073] Example 6:

[0074] This example utilizes the extractant trialkylphosphine oxide to enhance the transamination fermentation of recombinant strain E. coli LYJ201 to prepare furfurylamine. The process is carried out according to the following steps:

[0075] Step 1: inoculate the recombinant strain single colony on a solid LB plate into LB liquid medium, and add 1‰ chloramphenicol, 37℃, 200 rpm, cultivate for 8 h to obtain a primary seed solution;

[0076] Step 2: inoculate the primary seed solution into 250 mL fermentation shake flasks containing 75 mL M9 medium at a 2% (wt) inoculation amount, add 1‰ chloramphenicol when inoculating the primary seed solution, 180 rpm, 37℃, cultivate for 8 h to obtain a secondary seed solution;

[0077] Step 3: inoculate the secondary seed solution into a 5 L fermentation tank containing 1.5 L M9 medium at a 5% (wt) inoculation amount for fermentation, add 1‰ chloramphenicol when inoculating the seed solution, 500 rpm, 37℃, cultivate for 11 h;

[0078] Step 4: cultivate to OD 600 15, then add the inducer IPTG to a final concentration of 50 μM, 30℃, 500 rpm, induce expression for 3 h, then add the substrate furfural at a flow rate of 4.5 g / (L.h), add the amino donor (ammonium chloride) at a flow rate of 6 g / (L.h), add glucose at a flow rate of 12 g / (L.h), add the extractant trialkylphosphine oxide at a flow rate of 75 mL / h for 6 h after transamination for 4 h, continue the transamination reaction for 12 h to prepare furfurylamine. Centrifuge the sample, dilute the supernatant 50 times, then use HPLC to detect the furfurylamine concentration, which is 30.2 g / L.

[0079] Although the present application has been disclosed with reference to the preferred embodiments above, it is not intended to limit the present application, and any person skilled in the art can make various modifications and modifications without departing from the spirit and scope of the present application, therefore, the protection scope of the present application should be defined by the claims.

Claims

1. A process for the in situ extractive fermentation production of furfuryl amine, characterized in that, The method is carried out in the following steps: Step 1: inoculate the activated transaminase-producing recombinant engineering strain into a fermentation medium for culture; the recombinant engineering strain is E. coli LYJ201, and the recombinant engineering strain is obtained by knocking out DNA binding transcriptional dual regulator arcA and overexpressing transaminase gene OATA, alanine dehydrogenase gene ALD and T7 promoter connected carbon storage regulator CsrB gene based on E. coli BL21 (DE3) as a starting strain; the gene sequence of the DNA binding transcriptional dual regulator arcA is shown as SEQ ID NO. 4, the transaminase gene OATA is derived from Ochrobactrum anthropi, and the gene sequence is shown as SEQ ID NO. 3, the alanine dehydrogenase gene ALD is derived from Bacillus subtilis, and the Genebank ID is 936557, the sequence of the carbon storage regulator CsrB gene is shown as SEQ ID NO. 1, and the nucleotide sequence of the T7 promoter is shown as SEQ ID NO. 2; Step 2: after adding an inducer to induce fermentation, an external flow of a furfural, an amino donor and glucose is added for transamination reaction, and then an external flow of an extractant is added for extraction fermentation, the extractant can extract furfurylamine produced in situ in the fermentation broth; the induction fermentation refers to when OD 600 is 10~20, an inducer IPTG is added to a final concentration of 50~200 μM, and induction is carried out at 25~35℃, 100~1000 rpm for 2~6 h; the furfural flow rate is 0.5~5 g / (L·h), the amino donor is ammonium chloride, the flow rate is 0.5~6 g / (L·h), and the glucose flow rate is 1.5~15 g / (L·h); the extractant includes any one of trialkyl phosphine oxide, trioctyl phosphate, tributyl phosphate, n-octane and dodecane, the extractant is added at a volume fraction of 20%~50% based on the volume of the fermentation broth, and the flow rate is 0.03-0.18 V / h; the extraction fermentation culture conditions are 25~40℃, 100~1000 rpm, and 10~15 h. Step 3: after the fermentation is completed, the extractant is recovered, and furfurylamine is obtained by distillation under reduced pressure.

2. The in situ extraction fermentation production process of claim 1 wherein, The inoculation amount of the recombinant engineering strain in step 1 is 1% to 10% of the volume of the fermentation medium.

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