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Fermentative production of non-volatile microbial metabolism products in solid form

A non-volatile, metabolite technology, applied in the field of non-volatile microbial metabolites, which can solve complex problems, cannot be widely used, and is not optimized.

Inactive Publication Date: 2008-11-05
BASF AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0017] Therefore, although it is technically possible to process cassava as a starch source in a method equivalent to dry milling, this cassava-based method is still complex, unoptimized, and thus not widely applicable
To date, there has been no report on the use of grains as a starch source for the production of fine chemicals such as non-volatile microbial metabolites in a process equivalent to dry milling

Method used

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  • Fermentative production of non-volatile microbial metabolism products in solid form
  • Fermentative production of non-volatile microbial metabolism products in solid form
  • Fermentative production of non-volatile microbial metabolism products in solid form

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0236] a) Enzymatic starch liquefaction and saccharification

[0237] 500 grams of dry ground grits were suspended in 750 ml of water and finely ground again in a stirring mixer. The suspension was divided into four samples, Nos. 1 to 4, and each sample was treated with about 3 grams of thermostable alpha-amylase (Samples Nos. 1 and 2: Termamyl L; Samples Nos. 3 and 4: Spezyme). Samples No. 2 and No. 4 were then treated with approximately 7 g / l glucoamylase (Sample No. 2: Dextrozyme GA; Sample No. 4: Optidex). This gave pale yellow viscous samples, the solid contents of which were separated by centrifugation in each case, with a layer of hydrophobic solids floating above the clear liquid phase.

[0238] The clear supernatant of each sample obtained in this way was analyzed using HPLC in concentrated form and after 10-fold dilution, ignoring or taking into account the precipitate that had been centrifuged. When considering the precipitate, it is assumed that the dry matter co...

Embodiment 2

[0270] Using the corn meal hydrolyzate obtained according to Example II.1, with ATCC13032lysC described in WO 05 / 059144 fbr The strains were fermented analogously to example 1 b). Cells were incubated on sterile CM agar (composition see Table 4; 20 minutes at 121°C) for 48 hours at 30°C. Cells were then scraped from the plate and resuspended in saline. In each case, the optical density at 610 nm was achieved using the thus-prepared 610 A cell suspension with a value of 1 was used to inoculate 25 ml of medium 1 or 2 in a 250 ml Erlenmeyer flask (see Table 5). The samples were then incubated in a humid shaker (85% relative atmospheric humidity) at 200 rpm at 30° C. for 48 hours. The lysine concentration in the medium was determined by means of HPLC. In all cases, approximately equal amounts of lysine were produced.

[0271] The resulting lysine-containing fermentation broth was processed as described in Example 1c.2) to produce extrudates.

Embodiment 3

[0273] Corynebacterium glutamicum (ATCC13032lysC fbr ) in the shake flask test (shake flask 1+2) using the corn meal hydrolyzate obtained according to Example II.3a. Furthermore, wheat semolina hydrolyzate (shake flask 3+4) and rye semolina hydrolyzate (shake flask 5+6) prepared analogously to Example II.3 were used in parallel.

[0274] 3.1) Preparation of inoculum

[0275] Cells were streaked on sterilized CM+CaAc agar (composition: see Table 7; 20 minutes at 121°C) and then incubated at 30°C for 48 hours. Then inoculate on fresh plates and incubate overnight at 30°C. Cells were then scraped from the plate and resuspended in saline. In each case, the OD was achieved at the optical density value at 610 nm as prepared 610 A cell suspension with a value of 0.5 was inoculated in 23 ml of culture medium in a 250 ml Erlenmeyer shake flask (see Table 8).

[0276] Table 7: Composition of CM+CaAc agar plates

[0277] concentration

Element

10.0g / l

D-gl...

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Abstract

The invention relates to a method for production of at least one non-volatile microbial metabolism product in solid form by sugar-based microbial fermentation, wherein a microbial strain producing the required metabolism product is cultivated with a sugary liquid medium with a monosaccharide content of more than 20 wt. %, based on the total weight of the liquid medium, the volatile components of the fermentation brew are mostly removed, the sugary liquid medium being produced by: a1) milling a starch source selected from cereal grains, a2) liquefying the milled material in an aqueous liquid in the presence of at least one starch-digesting enzyme and subsequent saccharification using at least one saccharifying enzyme, wherein the liquefying is carried out with addition of a partial amount of the milled material to the aqueous fluid continuously or batch-wise. The invention further relates to a solid formulation obtained by the above method of a non-volatile microbial metabolism product and the use of such a solid formulation as additive or supplement to animal or human food or for textile, leather, cellulose, paper or surface treatments.

Description

field of invention [0001] The present invention relates to the fermentative production of non-volatile microbial metabolites in solid form by milling, liquefying and saccharifying a starch source selected from grains and by using the resulting sugar-containing broth for fermentation. Background technique [0002] Methods for the production of non-volatile microbial metabolites such as amino acids, vitamins and carotenoids by microbial fermentation are generally known. Depending on the process conditions, different carbon sources are used for this purpose. They range from pure sucrose to sugar beet and cane molasses, to so-called high-test molasses (sugarcane conversion molasses), to glucose from starch hydrolysates. In addition, acetic acid and ethanol are mentioned as auxiliary substrates (Pfefferle et al., Biotechnological Manufacture of Lysine. Advances in Biochemical Engineering / Biotechnology, Vol.79 (2003), 59- 112). [0003] Based on the above carbon sources, variou...

Claims

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Application Information

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IPC IPC(8): C12P19/14C12P13/08C12P13/14A23L7/104A23L35/00
CPCC12P13/08C12P7/46A23L1/105C12P13/10C12P19/14C12P13/14C12P13/02A23K1/007C12P13/12C12P13/24A23K10/12A23L7/104C12P13/04
Inventor M·蓬佩尤斯S·弗里尔M·洛沙伊德特O·策尔德尔M·博伊E·朔尔腾
Owner BASF AG
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