Method for Producing Glucosamine by Culturing Microorganism with Low-cost Medium

a microorganism and low-cost technology, applied in the direction of fertilization, etc., can solve the problems of affecting the purity of glucosamine, affecting the metabolism of cells in joints, and lowering the glucosamine in the human body, so as to reduce the cost of the medium

Inactive Publication Date: 2012-06-14
YUAN ZE UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The method achieves higher glucosamine concentrations and lower production costs compared to conventional techniques, with Aspergillus sp. producing up to 5.48 g / dm3 and Monascus pilosus producing glucosamine efficiently, while minimizing medium costs, thus addressing the limitations of existing methods.

Problems solved by technology

However, as age increased, the synthesis speed for glucosamine in the human body is lower than that of the decomposition speed of glucosamine, and consequently, the body and joints tend to be short of glucosamine, and further, the metabolism of cells in joints may be affected.
However, shrimp and crab carapaces obtained from different sources may affect the purity of glucosamine.
In addition, glucosamine produced form contaminated shrimp and crab carapaces may be toxic.
Unfortunately, since the regulatory mechanism involved was too complicated such that the detectable amount of glucosamine in the medium for E. coli became extremely low, only several milligram per liter.
Unfortunately, fast degradation of glucosamine in host cell, inhibition effect of glucosamine and degradation product thereof might hinder the increase of the glucosamine concentration [Deng, Ming-de, K. D. Severson, D. A. Grund, S. L. Wassink, R. P. Burlingame, A. Berry, J. A. Running, C. A. Kunesh, L. Song, T. A. Jerrell and R. A. Rosson, From Conceptto Process: Metabolic Engineering for Production of Glucosamine and N-Acetylglucosamine, Metabolic Engineering, 7(3), 201-214 (2005)].
Many Aspergillus fungi may produce secondary metabolites harmful to human body.
In view of the foregoing, conventional methods for producing glucosamine have still many disadvantages, and among the other, the production of glucosamine from microorganism fails to increase greatly as well as the cost of the medium is impossible to cut down, and consequently, are not well-designed and needs improvement urgently.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Test Medium

1. Experimental Strains

[0026]The invention discloses a method for producing glucosamine by fermenting a non-gene-transferred microorganism with changing medium. The microorganism used was one of the following: Monascus pilosus BCRC31527, with an accession number corresponding to ATCC 22080, and Aspergillus sp. BCRC31742, with an accession number corresponding to UPCC 3868. Both of the above-mentioned strains were purchased from Food Industry Development and Research Institute, Hsin-chu, Taiwan, ROC.

2. Medium

[0027]An appropriate medium was selected based the different characteristics of the above-mentioned two strains for carrying out the production of glucosamine. Media selected for each of the two strains were listed in Table 1.

TABLE 1Media for various glucosamine-producing strainsComponentsConcentrationStrainMediumin medium(g / L)MonascusRBA (pH 5)Rice bran25pilosusB-grade white25BCRC31527crystal sugarNH4Cl15Aspergillus sp.WP1 (pH 7)Superior white fineBCRC31742granulated ...

example 2

Shaking-Flask Fermentation Test

[0036]Various strains described in Example 1 was activated separately through three region streak plate culturing in PDA solid medium (Potato Dextrose Agar: 200 g / L Diced potatoes, 20 g / L Glucose, 15 g / L Agar) at 30° C. for 5 days. Then, single colony was picked up and placed in 200 cm3 sterilized PDB liquid medium (Potato Dextrose Broth: 20 g / L Diced potatoes, 4 g / L Glucose) contained in a 250-cm3 shaking-flask, followed by secondary activation through incubating in a thermostatic incubator at 30° C. and 200 rpm for 7 days.

[0037]Stains thus-activated was used to inoculate in a suitable medium (Table 1), and the pH of the medium was controlled as followed: pH of the medium used for microorganism Monascus pilosus BCRC 31527 was pH 5, pH of the medium used for microorganism Aspergillus sp. BCRC 31742 was pH 7. Thereafter they were incubated at a temperature of 30° C., and 200 rpm for 7 days. Samples were taken and the cell dry weight and yield of glucosa...

example 3

Fermentation Test in Fermenter

[0047]In this example, at first, Aspergillus sp. BCRC 31742 described in Example 1 was subjected to secondary activation culturing in accordance with the manner as in Example 2. Then, a fermentation test was carried out in a batchwise stirring fermenter as described below. The 100 cm3 thus-activated Aspergillus sp. BCRC 31742 liquor was inoculated in WP1 medium wothin the fermenter, and with a operation volume of 2 dm3, its was subjected to fermentation culture under optimal conditions obtained from shaking-flask experiment: pH 7, a temperature of 30° C., a rotational speed of 200 rpm, and with pure oxygen introduced through external lines to control the dissolved oxygen concentration at 10%.

[0048]After recovered from the fermenter, the fermentation liquor was filtered with suction to yield bacterial cake. Next, a sample of the bacteria cake was dried at 100° C., and the ratio of wet cell weight to dry cell weight was determined. Separately, wet cells w...

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PUM

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Abstract

A method for producing glucosamine with microorganism comprises of fermenting with a microorganism selected from the group consisting of Monascus pilosus and Aspergillus sp. in a novel low-cost medium, thereby enable it to produce glucosamine; wherein said medium is consisted of commercial Taiwan sugar, soy beam, rice bran and the like; wherein suitable condition for the fermentation is: 150˜300 rpm, pH 4˜pH 8, and 24° C.˜37° C.; wherein, after fermentation culturing, the fermentation liquor is filtered with suction to recover said microorganism biomass, said microorganism biomass is then subjected to steps of cell disruption, hydrochloric acid reaction, neutralization reaction and filtration, to obtain glucosamine produced by the microorganism.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]The present application is a divisional of U.S. patent application Ser. No. 12 / 407,171 filed on Mar. 19, 2009, which is incorporated by reference herein in its entirety.BACKGROUND OF THE INVENTION[0002]1. Field of the Invention[0003]The invention relates to a method for producing glucosamine by culturing microorganism with a novel low-cost medium.[0004]2. Description of the Prior Art[0005]Glucosamine is one of the constitutional ingredients for articular cartilage, which can provide nutrition for articular tissue, enhancing the ability of synovial fluid for recovering lubricating function, promote the regeneration of retrograded joints, and hence reduce effectively the pain generated from bone friction, as well as prevent the aggravation of arthritis condition. Glucosamine can be synthesized by human body itself. However, as age increased, the synthesis speed for glucosamine in the human body is lower than that of the decomposition speed ...

Claims

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Application Information

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Patent Type & AuthorityApplications(United States)
IPC IPC(8): C12P19/26
CPCC12P19/26
InventorWU, HO-SHINGCHANG, YU-FENWEI, YU-CHIAO
OwnerYUAN ZE UNIV