CS200884B3 - Process for producing exocellular glucanoic bipolymere produced by bacterial microorganism - Google Patents

Description

(54) A method for obtaining an exocellular glucan biopolymer produced by a bacterial microorganism

The present invention relates to a process for obtaining an exocellular glucan biopolymer produced by a bacterial microorganism on nutrient media containing carbon, nitrogen and biologically active substances by submersible sterile fermentation and isolating its own polysaccharide biopolymer for use in the geological, food and pharmaceutical industries,

Based on the present knowledge, which relates mostly to the glucan type polysaccharide produced by Leuconostoc cultures and the glucomannan type produced, for example, by Xanthomonas or Arthrobacter cultures, the fermentation of the glucan type produced by the aerobic production strain of Achromobacter delicatulus has not been described in the literature. For example, we know from the literature that the polysaccharide of the genus Xanthomonas (U.S. Pat. No. 3,485,719, 3,096,293, or 3,328,262) is composed of glucose and mannose, but whose fermentation viscosity is lower and the fermentation is more susceptible to secondary contamination.

SUMMARY OF THE INVENTION The present invention provides a process for obtaining a novel exocellular biopolymer produced by the bacterial microorganism Achromobacter delicatulus (Jordan, 1890) (Bergey et al. 1923) No. 39, which is described in U.S. Pat. No. 172 624, which is based on the inoculation of a 24-hour culture of bacteria

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200 004.2 Aohromobaoter delicatulus (Jordan, 1890) (Bergey et al. 1923) 8,39 are fermented into the culture medium for 48 to 72 hours at 28 to 30 ° C at pH 6.8 to 7 * 2 in three stages, wherein an additional sugar source is added at each stage, for example preferably raw sugar, whereupon the biopolymer is isolated after ethanol fermentation or by directly drying the unfermented soil by spraying or hydrolysis and fractionating to produce a biopolymer of the desired molecular weight. It is an aerobic production strain whose glucane-type polysaccharide gives the following monomeric sugars after hydrolysis: D-ribose, D-xylose, D-arabinose, D-mannose, D-fructose, and D-galactose 7 in amounts from 0.3 to 3 % wt. and Ζ-D-glucose and β-D-glucose in amounts of 85 to 87% by weight.

The polysaccharide obtained by the process of the invention has a higher viscosity of the fermented soil than the prior art polysaccharides fermented from other microorganisms and is less prone to contamination during fermentation. The homogeneity of this polymer is documented in its elution diagram in Fig. 1, its infrared absorption spectrum in the diagram in Fig. 2, and the composition of the polysaoharide as a mixture of trimethylsilyl monosaoharide derivatives in Fig. 3.

In contrast to the previously described glucane-type polysaccharides fermented predominantly anaerobically (for example, Leuconostoc culture), a multi-step industrial aerobic cultivation of polysaccharides of said properties has not been described by the production strain Aohromobacter delicatulus (Jordan, 1890) (Bergey et al. 1923) No. 39.

The following is an example of a method for obtaining a biopolymer according to the invention.

Example

First, prepare the inoculum in a pre-seed tank with a volume of 400 l filling 200 l of the medium using these raw materials (culture medium)

raw sugar 3 wt. oorn-steep (CSL) 0.2 wt. K ₂ P ° ₄ 0.75 wt. NaCl 0.01 wt. MgSO ₄ . 7 HgO 0.02% wt. kno ₃ 0.04% to 0.4% wt soybean oil 0.1 wt. yeast autolysate 100% in aqueous environment (tap water) 0.2 wt.

The pH was adjusted to 6.8 with 40% NaOH.

The inoculation tank was inoculated with 10 1 inoculum of Aohromobaoter delicatulus (Jordan, 1890) (Bergey et al. 1923) # 39, 24 hours old.

Cultivate for 24 to 26 hours at 28 to 30 ° C with stirring and aeration under sterile conditions (1st stage).

Then, the contents of the pre-seed tank were transferred to a 3500 L seed tank with a 2000 L feed (2nd stage). Fermentation lasted 48 hours at 28-30 ° C. The pH was maintained at 6.8 under the same conditions as in the first step. At 48 hours, the contents of the seeding tank were transferred to a fermentation tank (3rd stage, 35 nP content, loading 20 m plnění).

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At 78 hours, a 1.5% weight / weight portion of raw sugar was added to increase yield. (At 122 hours of fermentation, the viscosity of the fermentation fluid measured with a rotary viscometer at 12 rpm was 26150 oP). Fermentation was complete at 136 hours. Then, half the volume was treated with ethanol precipitation, and the polymer obtained was dried in a hot air oven. The 1% solution of the crude powder biopolymer thus obtained had a value of 640 oP. 308 kg of a 86.9% biopolymer were obtained.

The other half of the tank was treated with an Anhydro spray dryer at an inlet temperature of 175-188 ° C and an outlet temperature of 108-109 ° C. A total of 400 kg of a yellow powder of 97.1% by weight was obtained. solids containing 61 to 68% pure biopolymer.

The biopolymers obtained by the process of the invention have the excellent rheological and structural mechanical properties required for polymer drilling when applied to deep wells (e.g., diesel) in the preparation of osmotic drills.

SUBJECT OF THE INVENTION

Claims (1)

SUBJECT OF THE INVENTION A method for obtaining an exocellular glucan biopolymer produced by the bacterial microorganism Achromobacter delicatulus (Jordan, 1890) (Bergey et al. 1923) No. 39, which is described in the description of the invention to U.S. Pat. No. 172,624, characterized in that after inoculation of a 24-hour culture of Achromobacter delicatulus (Jordan, 1890) (Bergey et al. 1923) No. 39 in the culture medium, it is fermented for 48 to 72 hours at 28 to 30 [deg.] C at pH 6.8-7.2 in three stages, with an additional sugar source, preferably raw sugar, added at each stage, after which the biopolymer is isolated after ethanol fermentation or by spray drying or hydrolyzing the fermented soil directly, and the biopolymer is prepared by fractionation of the desired molecular weight. 3 drawings