Isolation and amplification of the phy gene coding phytase from bacillus sp.

Abstract

Poultry are unable to metabolize phytic acid in feed due to no enzyme to digest and degrade it. Therefore, phytic acid becomes an anti-nutritional substance in poultry feed which can result in decreased feed efficiency, decreased nutrient intake which gave a negative impact on health and production. The addition of phytase in the poultry ration is necessary to degrade phytic acid. However, the availability of phytase in Indonesia is still limited because technology in Indonesia is still inadequate to produce phytase, so Indonesia cannot produce it itself and still imports phytase at quite high prices. This study aims to isolate and amplify the phy gene encoding phytase derived from native microbe"s genus Bacillus from Indonesia. The method used in present study was to isolate the genome of Bacillus using cell lysis method, then amplify the genome using polymerase chain reaction (PCR) method. The results of this study were that genome isolation results from Bacillus gene sources were obtained that the purity of A260/A280 from B. sp 6, B. sp 7, and B. licheniformis was 1.86 μg/mL, 1.95 μg/mL and 1.86 μg/mL, respectively. Furthermore, the present study showed that 0.4 μM/μL primer concentration was able to amplify the phy gene. 200-400 ng/μL DNA template concentration can produce optimal DNA bands during amplification of the phy gene in Bacillus sp. In conclusion, amplification was successfully carried out on B. sp and B. licheniformis. The length of the phy gene in B. sp is 1149 bp and the phy gene in B. licheniformis is 1146 bp. The genus Bacillus native to Indonesia has the potential to be continued as a source of phytase producing genes.

Keywords

• amplification
• bacillus
• isolation
• pcr
• phytase