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Fermentation Optimization and Production in Pilot Scale of Microbial Transglutaminase Using Streptomyces Hygroscopicus WSH03-13

Author: ZhangChengJie
Tutor: ChenJian
School: Jiangnan University
Course: Fermentation Engineering
Keywords: Transglutaminase Identification of bacteria Streptomyces hygroscopicus Fermentation process optimization Pilot production Hydroxylamine Inducer
CLC: TQ925
Type: Master's thesis
Year: 2005
Downloads: 243
Quote: 3
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Abstract


Microbial transglutaminase (protein - glutamic acid-γ transglutaminase, EC 2.3.2.13, MTG) can catalyze in vitro as a variety of proteins, protein and amino acid crosslinking reaction and improve various proteins functional properties, and thus in the food, cosmetics, pharmaceutical and other industries have broad application prospects. This study first a transglutaminase producing strains WSH03-13 were identified, identified as: Streptomyces (Streptomyces hygroscopicus) WSH03-13. With this as a starting strain strains, through the choice of carbon and nitrogen sources, optimizing the fermentation medium; industrialization as the goal for the fermentative production of transglutaminase from 7L to 150L small test pilot scale amplification test; visits the inducer for Streptomyces hygroscopicus WSH03-13 enzyme production. The main contents are as follows: 1. Strain WSH03-13 and 16S rDNA sequences of Streptomyces (Streptomyces sp.) Of 16S rDNA sequence homology. According to the physiological and biochemical characteristics of the strain as well as some 16S rDNA sequence alignment, strain WSH03-13 as Streptomyces (Streptomyces hygroscopicus), its taxonomic status as: Bacteria; Actinobacteria; Actinobacteridae; Actinomycetales; Streptomycineae; Streptomycetaceae; Streptomyces. 2 pairs of strains WSH03-13 fermentation medium was optimized. Through experiments have selected the appropriate carbon and nitrogen source and optimizes complex combination of carbon and nitrogen sources to determine the fermentation medium formulations. 3 In the optimized fermentation conditions were 7 L fermenter small test. 1.25 vvm aeration volume and stirring rate of 400 rpm, so that the dissolved oxygen in the fermentation broth was maintained at 30 to 25% level, to meet the Streptomyces hygroscopicus WSH03-13 fermentation MTG demand for oxygen. 7 L fermentor highest enzyme Activity reaches 4.48 u / ml. 7 L fermenter small trial found that oxygen is a key factor in the fermentation process. 4 compares the 30 L and 150 L fermentor test results. In small magnification (4 times) in the case, 30 L fermenter can completely reference 7 L pilot fermenter operating process conditions to achieve a similar level of activity. In the 150 L fermentor (20x), due to pot size and equipment conditions change, making the process conditions need to make appropriate adjustments. Oxygen around the key factors of the fermentation process, the ventilation volume increase to 1.5 vvm, agitation rate selection at 250 rpm. 30 L tank test, MTG highest activity of 4.26 u / ml; 150 L tank test, MTG highest activity reached 4.20 u / ml, close to the small scale level. 5 of adding hydroxylamine right Streptomyces hygroscopicus WSH03-13 synthetic transglutaminase enzymes. The results showed that the addition of hydroxylamine to induce the synthesis of transglutaminase. In the optimum dosage and hydroxylamine optimum addition time was 0.5 mM and 38 h, the end of fermentation, MTG highest activity 5.3 u / ml, was increased by 32.5%.

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