# US 11,111,512 B2 · Method for producing 5-hydroxytryptophan

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- Number: US 11,111,512 B2
- Document ID: US11111512B2
- Status: Granted
- Kind: B2
- Application number: 16/311,381
- Filed: 2017-06-05
- Published: 2020-06-25
- Granted: 2021-09-07
- Earliest claimed: 2017-06-05
- Assignees: Blrh Biotech Co.; Nanjing Shoubai Biotech Co.
- Applicants: Blrh Biotech Co.; Nanjing Shoubai Biotech Co.
- Inventors: Yunxian Zhao
- Web page: https://patentgrep.com/patent/US11111512B2

## CPC

- C12P 13/22: Tryptophan; Tyrosine; Phenylalanine; 3,4-Dihydroxyphenylalanine
- C12P 13/227: {Tryptophan}
- C12N 1/20: Bacteria; Culture media therefor
- C12N 1/205: Bacterial isolates
- C12P 7/08: produced as by-product or from waste or cellulosic material substrate
- C12R 2001/10: Bacillus licheniformis
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS

## Abstract

This invention involves to Bacillus licheniformis JSC-69 for producing the 5-HTP, deposited as CGMCC NO: 13533; and a method for the producing 5-HTP using Bacillus licheniformis. Bacillus licheniformis JSC-69 said in this invention produces 5-HTP using tryptophan as the substrate, and the transformation efficiency is 95%˜100%.

## Claims

### Claim 1

A method for producing 5-HTP, by Bacillus licheniformis JSC-69 deposited under accession number CGMCC 13533, said method comprising the following steps:
Step a) culturing of Bacillus licheniformis said Bacillus licheniformis JSC-69;
Step b) adding tryptophan to said culture of step a) and transforming the tryptophan to 5-HTP by said Bacillus licheniformis JSC-69;
Step c) collecting the 5-HTP.

### Claim 2 (depends on 1)

The method of claim 1, wherein the culturing step comprising any one or a combination of slant culturing, shake flask culturing, seed tank culturing and fermenter culturing.

### Claim 3 (depends on 2)

The method of claim 2, wherein the fermenter culturing is carried out under 30-35° C. and pH 6.5-7.0 for 30-50 h.

### Claim 4 (depends on 2)

The method of claim 2, wherein the medium of fermenter culturing comprises sucrose, bean pulp, (NH4)2SO4, K2HPO4.3H2O and KH2PO4.

### Claim 5 (depends on 1)

The method of claim 1, wherein the tryptophan concentration in Step b is 0.2-10 g/L.

### Claim 6 (depends on 1)

The method of claim 1, wherein the transforming in Step b is carried out under 25-45° C. and pH 6-8 for 40-60 h.

### Claim 7 (depends on 1)

The method of claim 1, wherein the collecting described in Step c includes the following method: centrifuging the transformed product obtained in Step b and collecting the supernatant, which is the product 5-HTP.

### Claim 8 (depends on 1)

The method of claim 1, further comprising Step d, that is purifying the product 5-HTP, the “purifying” process includes solid-liquid separation, cation exchange resin adsorption separation, concentration, ethanol dissolution and distillation steps in order of sequence.