Rapid sanitary and technological control of culturing the producers of amylolytic enzymes.

Abstract

It has been established that the biotechnological process of culturing bacillary microbial producers of amylolytic enzymes can be express-controlled by determining their ATP bioluminescence. The advantages of the method have been shown. The analytical review of producers of hydrolytic enzymes has made it clear how practical it is to use bacillary microorganisms for targeted secretion of amylolytic enzymes in biotechnological production. After monitoring bacillary microorganisms, it has been found advisable to choose Bacillus subtilis ATCC 6633 as the working culture due to its time of production of amylolytic enzymes and its biosynthetic activity. Reasons have been given for using the rapid ATP control method, which is based on the principle of bioluminescence. Different growth media have been compared and evaluated in order to intensify the quantitative biosynthetic activity of the microbial culture in the technological process of culturing bacillary microorganisms. The experiments have proved that growth media can be modified by introducing a number of carbohydrate–protein substrates as inducers of amylolytic complex gene expression. The latter manifests itself in the amylolytic activity accelerated by 12–24 hours, and causes an increase in the number of microorganisms (1.87–3.99 times as many as in the reference culture). Two methods of control (rapid bioluminescent and classic microbiological) have been used for correlative determination of the quantitative growth of Bacillus subtilis cells. Mathematical straight-line correlations have been obtained in a semilogarithmic system for the number of cells of the bacillary producer of the amylolytic enzyme complex. These correlations allow carrying out rapid control in a production environment. Along with the traditional rapid sanitary control in biotechnological production, which includes controlling the contamination of the equipment, personnel’s hands, and water, it has been suggested to perform proprietary technological express control of amylolytic enzyme biosynthesis using the culture Bacillus subtilis ATCC 6633

Keywords

• bacillary microorganisms
• bacillus subtilis
• atp (adenosine triphosphate)
• bioluminescence
• amylolytic enzyme complex
• sanitary and technological control