JPS5934894A - Preparation of l-proline by fermentation - Google Patents

Abstract

PURPOSE:To prepare L-proline, by culturing microorganism belonging to Bacillus genus using milk whey, soybean whey, etc. as a lactose or galactose source. CONSTITUTION:A microbial strain belonging to Bacillus genus and capable of producing proline assimilating galactose or lactose, e.g. Bacillus subtilis AJ3010 (FERM-P No.6675), is inoculated in a medium containing lactose or galactose as the carbon source, and cultured under aerobic condition at 27-37 deg.C and 5.5- 7.5pH for 1-4 days. L-proline produced and accumulated in the medium is separated therefrom.

Description

[Detailed description of the invention] The present invention relates to a method for producing monoproline by a fermentation method.

■ In the production method of 7-broline (hereinafter simply referred to as proline) by fermentation, there is a method using a proline-producing mutant strain requiring amino acids derived from L-glutamic acid-producing bacteria, a method using a proline-producing mutant strain of Bacillus spp. , a method using a histidine-, L-methionine-, or L-leucine-requiring mutant strain of the genus Ennerlichia (see Japanese Patent Publication No. 26911/1973), etc. are known. Glucose is used as a carbon source in these proline fermentation methods.

The inventors of the present invention have repeatedly conducted various studies on a method of fermenting and producing proline using milk whey, which is inexpensive and has little utility value, as a raw material. As a result, the inventors have found that lactose, the main component of milk whey, is present in bacteria belonging to the genus Bacillus isolated from nature. discovered that there are microorganisms that have the ability to assimilate proline.

The present invention was completed based on this discovery. The present invention will be explained below.

The microorganism used in the method of the present invention is a microorganism that has the ability to assimilate galactose or lactose to produce proline, and a representative example is Bacillus subtilis.
AJ3010FERM-P 667 is mentioned.

The medium used for culturing such Bacillus species having proline-producing ability is not particularly different except that it contains lactose or galactose as a carbon source. As a source of lactose and galactose, milk whey, soybean whey, etc. containing these can be used. In addition to these carbon sources, the culture medium may also contain supplementary carbon sources such as gluten and 7-ucrose. In addition to carbon sources, ordinary nitrogen sources such as ammonium ions, ammonia gas, and ammonia water, non-deposited ions such as phosphate ions, magne/lamb ions, and potassium ions, and organic micronutrients such as vitamins and amino acids are also used as necessary. Contained in the medium.

Cultivation is carried out under aerobic conditions. During cultivation, it is desirable to maintain the culture temperature at an appropriate temperature within the range of 27 to 37°C and the medium pH at an appropriate pH within the range of 5.5 to 7.5. Thus, if the culture is continued for 44 days, bourin will be produced and accumulated in the medium.

Proline accumulated in the medium can be collected by conventional methods.

Examples will be described below.

Example 1 Galactose 8? /dl, N H4C11,0 you.

Kca O,2q7ti7! , KH2PO40,Ir
J, )#SO4・7H200,04? /di,
Casamino acids 0, 4 fl/dl, FeSO4・7 [
01H9/d7! , MnSO4・4H, OI mg
/dl and adjusted to pH 7.0 at 5oo+M
! One 20ml flask was placed and sterilized. One platinum loop of Bacillus subtilis AJ 3010 was planted in this,
The cells were cultured at 31°C for 48 hours. In addition, during cultivation, 459/
d7! of urea water was added in 0.3 ml portions to prevent the pH from decreasing.

The amount of Shiichiproline accumulated at the end of the culture was 120 m
g/de.

Example 2 Acid-precipitated milk whey with the following composition was used as lactose.4? /
dl, (NI-14)2 Soi 2.5? /
d & KH, PO i 0.2? /dl.

MgSO4・7H200,1? /di 1 yeast extract 0-05 ty'dl.

FeSO41H,,01m9/de, MnSO4・
4H201M9/di and Cal/Rum carbonate 2.5? /d
/, (separately added), a medium adjusted to pH 7.2 with KOH was prepared, placed in 20ml each of 500ml flasks, and sterilized. In this, Bacillus subtilis AJ3
0101 platinum loops were planted and cultured at 31°C for 72 hours.

The amount of proline accumulated at the end of the culture was 1101
19/d7! Met.

In addition, the ingredients of acid precipitated milk whey are 93.9% water and 4% lactose.
．． 5%, total solids 6.1%, nitrogen compounds 0.80%, ash 0.7096, and fat 0.1%.

Patent applicant: Ajinomoto Co., Inc.

Claims (1)

[Claims] A microorganism belonging to the genus Bacillus that has the ability to produce L-proline from lactose or lacto-X is cultured in a liquid medium containing lactose or galactose as a carbon source, and L-proline is produced in the culture solution. Seshime,
A method for producing proline by a fermentation method, which comprises collecting the proline.