การปรับปรุงผลผลิตของกระบวนการหมักสปอร์ของ Bacillus subtilis TISTR 001 เพื่อเป็นโพรไบโอติก

Productivity Improvement of Bacillus subtilis TISTR 001 Spore Fermentation for Probiotic

Name: จันทร์จีรา อยู่คง

Organization : มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี

keyword: Bacillus subtilis TISTR 001

; Response surface methodology

; Batch cultivation

; Fed-batch cultivation

Abstract: The use of Bacillus subtilis as probiotics for feed supplement in farming to promote forming bacteria which grow very fast. It can be produced in large-scale production. The objective of this study is to achieve both of high total cell concentration and spore concentration by media optimization and process improvement of spore fermentation. B. subtilis TISTR 001 was tested for the ability of microbial inhibition to some ofpathogenic bacteria (Staphylococcus aureus, Staphylococcus aureus LTH, Salmonella typhi and Salmonella anatum SO86105) by well diffusion and paper-disc assay. The result showed that undiluted supernatant of B. subtilis TISTR 001 did not inhibit these pathogenic bacteria. Inexpensive agricultural by-products such as molasses and defatted soybean meal can be used effectively in culture media. The response surface methodology (RSM) was applied for media optimization to get high spore concentration. A Central Composite design (CCD) was applied to study the combined effect of three factors (molasses, CaCl2.2H2O and MnSO4.H2O). The results showed that the optimized concentration of molasses (reducing sugar equivalent), CaCl2.2H2O and MnSO4.H2O were 19.85 g/L, 0.35 g/L and 0.15 g/L, respectively and the spore production was 5.50 x 109 spores/mL. However, in 2-L fermentor, the high concentration of molasses caused a foaming problem. The another optimization was performed in shake flask culture. Medium containing 20 g/L defatted soybean meal, 3 g/L molasses and 0.5 g/L K2HPO4 was suitable for spore production and resulted in a spore concentration of 1.30 x 109 spores/mL. Therefore, the scale-up of spore production was carried out with this medium. The batch cultivation of B. subtilis TISTR 001 in 2-L fermentor obtained a spore concentration of 1.78 x 109 spores/mL at 48 h, whereas the spore concentration in 1,500-L fermentor was 2.29 x 109 spores/mL at 47 h. In 75-L fed-batch cultivation of B. subtilis TISTR 001 with the same medium increased molasses to 5 g/L, the spore concentration was 5.73 x 109 spores/mL.

มหาวิทยาลัยเทคโนโลยีพระจอมเกล้าธนบุรี

Address: กรุงเทพมหานคร (Bangkok)

Email: info.lib@mail.kmutt.ac.th

Name: Phenjun Mekvichitsaeng

Role: Supervisor

Created: 2546

Issued: 2005-10-12

Modified: 2006-05-09

วิทยานิพนธ์/Thesis

ISBN: 9741833113

CallNumber: BIT480

tha

DegreeName: Master of Science

Level: Master's Degree

Descipline: Biotechnology

Grantor: King Mongkut's University of Technology Thonburi

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