JPS596894A - Preparation of raw material for fermentation - Google Patents

Abstract

PURPOSE:To prepare a raw material for fermentation of quality to improve the fermentation yield without containing impurities economically with a high efficiency, by converting sucrose in molasses into glucose and fructose, and carrying out the chromatography of the resultant product. CONSTITUTION:Invertase or an acid, e.g. hydrochloric acid or sulfuric acid, is reacted with cane molasses or beet molasses, etc. to convert the sucrose in the molasses into glucose or fructose, and the resultant product is then subjected to the chromatography with a cation type exchange resin to collect a fraction containing the glucose and fructose.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing fermentation feedstock from molasses.

Molasses is a by-product produced in the sugar industry, such as sucrose or beet sugar, and is mainly used in shakes containing p-Δ-Y.
However, since it contains a large amount of contaminants in addition to sugar, it is not possible to economically separate the sugar, and it is currently used as a raw material for glutamic acid fermentation or alcohol fermentation. However, even when used as a fermentation raw material, a large amount of impurities, especially colored substances, are transferred to the fermentation liquid as is, making it difficult to separate and purify the fermented product, and also from the perspective of waste liquid treatment. This also poses a large cost burden. Furthermore, when cane molasses is used not only for the removal of impurities but also as a raw material for fermentation, the fermentation yield is low, and in this respect it cannot be said to be satisfactory.

Therefore, the present inventors have conducted various studies with the aim of developing an economical and efficient method for producing fermentation raw materials of quality that improves the fermentation yield from molasses. After converting sucrose to glucose and fructose by the action of invertase or an acid, invert sugar and impurities can be efficiently separated by chromatography using a cation exchange resin, and that the invert sugar and impurities can be efficiently separated. By using a sugar solution (invert sugar solution), the yield of amino acid fermentation can be dramatically improved.
They also discovered that fermentation products can be separated and purified extremely easily, leading to the completion of the present invention. That is, the present invention involves converting sucrose into glucose and 7 lactose by using molasses invertase or an acid, and then performing chromatography using a cation exchange resin to separate invert sugar. The molasses used in the present invention is cane molasses, beet molasses, etc. First, molasses containing yucrose, which is molasses, is converted into glucose and fructose with invertase or acid. This is a complete conversion.

As invertase, commercially available enzymes as well as processed yeast cells having invertase activity can be used.

The yeasts used are those of the genus Sameromyces, Candida, Mitstorula, Debaryomyces, Vichia, Hansenula, and L-rulobcis, and these yeasts are mixed with molasses, glucose, valve manufacturing waste, soybean whey,
Yeast cells obtained by culturing in a medium containing food factory wastewater, fruit juice, etc. are used. Processed yeast cells include dried yeast cells (baker's yeast, brewer's waste liquid yeast), residues obtained by extracting useful components such as yeast extract, RNA, and glutathione from yeast cells, and autolysis. The resulting bacterial cell residue, physically crushed bacterial cells, and bacterial cells treated with lysozyme, toluene, and a surfactant are used.

Before the invertase in the molasses is converted, add the molasses to an appropriate concentration, e.g.
The enzyme reaction may be carried out by diluting oy/deT, adding the two invertase sources, and holding at 20 to 60 tll' for 5 to 20 hours.

The conversion of tsucrose with acid is carried out according to the known method of adding hydrochloric acid or sulfuric acid to molasses to adjust the pH of the molasses from 1.0 to 4.01, and heating it to a temperature of 80 to 110 tl. . After acid hydrolysis, add an alkali such as sodium hydroxide to neutralize. If a precipitate is produced in the acid decomposition-neutralization step, it is desirable to remove the precipitate, which can further enhance the purification effect.

The cation exchange resins used in the present invention include strong acidic cation exchange resins such as Hama Amberlite I R-120, Dowenx-50° Diaion 5K-IBs, Amberly 1-IRC-60, Amberlite XE-80,
Use one cation type such as Diamond ion WK-11 or another cation type such as Ca type.

To perform chromatography using these ion exchange resins, the cationic ion exchanger described above is packed in a column of an appropriate size, and the salts are eluted from the packed column, followed by glucose and fructose. be done. After the chromatography is completed, sugar and impurities can be efficiently separated by collecting invert sugar, that is, a fraction containing glucose and fructose.

On the other hand, when no conversion treatment is performed, there is almost no adhesion at the sucrose elution position and the colored substance elution position, so sucrose cannot be separated from impurities.

The sugar thus separated is desirable as a raw material for fermentation, and is used as a raw material for amino acid fermentation including glutamic acid fermentation.

5- The trowel of Example 1 will be explained below.

Example 1 Add 1 equivalent of water to 1 cane molasses, strain 1 ml of commercially available biocon as a coinvertase source (dry yeast cell wall sample manufactured by Biocon), and add 1.2 of molasses of sugar.
It was added at a rate of 1 g and kept at 55 ml for 10 hours to perform an enzyme reaction. Molasses treated with invertase as described above 1. A column (+, 5
Supply 1 x 30cnn) and elute with water. Add the eluate to 2. The solution was collected in 0ml portions, and each fraction was analyzed using high-performance liquid chromatography. The results are shown in FIG. As a control, the same chromatography was performed using 1 volume of Cane molasses diluted 2:1 with water without enzyme treatment.

The elution pattern is shown in FIG. In FIGS. 1 and 2, the vertical axis shows the stain density (v/dl), and the horizontal axis shows the fraction number. Further, impurities, glucose and fructose are expressed as -.-1- to 1 and hero - - -, respectively.

After the chromatography was completed, the sugar fraction was collected and 94 units of sugar were recovered. The impurity removal rate (removal rate of impurities other than sugar) of this sugar solution was 61. The impurity removal rate of this 1 sample versus the 1-1 control without invertase treatment was 17%, indicating that the separation performance could be dramatically improved by 1 point compared to 1 invertase treatment.

Example 2 Add ``Biocon'' to cane molasses diluted 2:1 with water
was added, and an enzymatic reaction was carried out in the same manner as in Example 1. 100 ml of invertase-treated molasses was transferred to a Ca-type cation exchange tree l1lif (Diaion SK-l B S
) 5. A column packed with OL was supplied and eluted with water. The eluate was analyzed by high-performance liquid chromatography, and the fraction containing glucose and fructose was collected to recover 95 units of sugar.

The impurity removal rate of this sugar solution was 59%. On the other hand, one cane molasses (only diluted 2-fold with water) that had not been subjected to invertase treatment was subjected to the same column coumadography, and 95 sugars were recovered. The impurity removal rate of this sugar solution was 14.

This chromatography was repeated one time, and the separated sugar solution was concentrated to a sugar concentration (sucrose equivalent) of 50.
A glutamic acid fermentation medium of 300 ml was prepared by mixing 50 ml of the sugar solution with 250 ml of a salt solution having the composition shown in Table 1.

Table 1 Composition of salt solution Content KW, PO42,09 MgSO4' 7H20Q,5 //FeSO4・7
H, 010mg Mn 504 ・4H20] 0 Thiamine hydrochloride 200 μ? Soybean protein hydrolysis solution (TN4r/dg) Ot
The mixture was poured into a fermenter and sterilized by heating using a 115 trr trowel for 10 minutes. This is a pre-cultured Brevibacterium.
Lactofermentum ATCC13869 was inoculated and 3
], 5trt trowel pH, ammonia gas 1 trowel 7.8t
The culture was carried out under aeration and stirring while maintaining the temperature. During culture, when the sugar concentration in terms of sucrose in the medium was below 3, the sugar solution used was added little by little to adjust the sugar concentration to 2-4% and cultured for 36 hours.

The amount of sugar solution added was the same for each experimental group at 80+++Jt. During the culture, when the predetermined amount of bacteria reached 1, the surfactant "Tween 60" was added in an amount of 0.6% t to t of the medium. Table 2 shows the amount of L-glutamic acid accumulated in the culture solution and the sugar yield. This is shown.

9- Table 2 I7-Glutamic acid fermentation yield No No ? , 52
46.4 No Yes 8.10
60.0 Yes No 7.9 B
48.0 Yes Yes
8.60 52.5-10- Example 3 Cane molasses was diluted twice with water and sulfuric acid was added to adjust the pH.
After adjusting the temperature to 25, the mixture was heated at 115r for 15 minutes to convert sucrose into glucose and fructose. This was neutralized by adding sodium hydroxide (pH
6,5) Centrifuge to remove the precipitate 1~. This acid decomposition solution was applied to an ion exchange column in the same manner as in Example 2.
7 tographies were performed. The sugar-containing fractions were then collected, the sugar was separated, the recovery rate was 95%, the impurity removal rate was 60 g), and the sugar solution was concentrated under reduced pressure to obtain a sugar solution with a sugar concentration (calculated as sugar) of 50 g.

When L-glutamic acid fermentation was carried out using this sugar solution in the same manner as in Example 2, the accumulated amount of L-glutamic acid was 8.49 v/di, and the yield based on sugar was 52.4%.

[Brief explanation of the drawing]

FIG. 1 shows a chromatoduran made from diamond ion 5KIBr from cane molasses treated with invertase, and FIG. 2 shows a chromatoduran made from diamond ion 5KIBr from cane molasses that was diluted twice with water. Patent applicant: Ajinomoto Co., Inc.

Claims (1)

[Claims] After converting molasses into sucrose glucose and fructose by acting on invertase or acid,
A method for producing a fermented raw material, which comprises performing P mudgraphy using a cation exchange resin to separate invert sugar.