COMPARATIVE ANALYSIS OF ALPHA AMYLASE AND GLUTAMINASE IN ASPERGILLUS ORYZAE ON FERMENTATION STARTER OF SOY SAUCE

• Main Author: Ulul Azmi, Adhigka
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/64099
• Institution: Institut Teknologi Bandung

Soy sauce is a common condiment found in cuisines in East Asian countries. In Southeast Asia, sweet soy sauce, such as that made in Indonesia and beyond, is more common than soy sauce. Originally, soy sauce was made in China in 160 BC through solid fermentation of soybeans which produced the liquid that is now known as soy sauce. When the Chinese introduced Buddhism to Japan, the knowledge of making soy sauce was brought along. Making soy sauce using Aspergillus oryzae as a mold for fermenting soy sauce. A. oryzae which is naturally found in soil cannot do sexual crossbreeding, and over time there was domestication of A. flavus which led to the emergence of A. oryzae which can be used in the production of soy sauce. Of the various enzymes that play a role in the transformation of organic matter in soybeans, alpha amylase and glutaminase enzymes play a key role in the production of compounds that form the character of the resulting soy sauce. This study aims to determine the differences between A. oryzae that have not been domesticated and those that have been domesticated using a phylogenetic analysis approach. Analysis was performed using Multiple Sequence Alignment (MSA), protein domain annotation, and phylogenetic analysis. Samples or reference data, taken from the NCBI gene bank and uniprot, are representative strains in the production of soy sauce. Fifteen samples for alpha amylase analysis consisted of 4 strains isolated from soy sauce, 1 strain from soju, and 6 strains from soil. Meanwhile, 8 samples for glutaminase analysis consisted of 3 strains isolated from soy sauce, 1 strain of soju isolate, and 4 strains of soil isolate. The amino acid sequence of alpha amylase shows the informative region on the order of 26-395. The amino acid sequence of glutaminase shows the informative region on the order of 341-512. The phylogenetic tree by utilizing these two enzymes significantly succeeded in differentiating isolates from soy sauce, from isolates obtained from soil. A phylogenetic tree using data on nucleotide variations in the area was able to distinguish the two groups. The use of various methods of constructing phylogenetic trees, such as maximum parismony, UPGMA, neighbor joining, and maximum likelihood, shows the same branching/grouping with a large bootstrap value of 79-100. As for the phylogenetic tree generated by the maximum likelihood method, it produces branching that still does not fully distinguish the two groups. From all the analyzes carried out, genetic information on the catalytic side of alpha amylase and glutaminase can classify the strains used in the production of soy sauce, and show the presence of branching caused by domestication in the manufacture of soy sauce.