Analisis gen-gen enzim pemecah karbohidrat isolat SU-KC1a = Analysis of genes encoding carbohydrate degrading enzymes in SU-KC1a

Kindangen, Wulan (2021) Analisis gen-gen enzim pemecah karbohidrat isolat SU-KC1a = Analysis of genes encoding carbohydrate degrading enzymes in SU-KC1a. Bachelor thesis, Universitas Pelita Harapan.

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Abstract

Pada ASI dapat ditemukan banyak bakteri jenis probiotik, salah satunya bisa berupa Lactiplantibacillus plantarum (L. plantarum). Diketahui bahwa L. plantarum dapat memfermentasi berbagai karbohidrat. Tujuan dari penelitian ini adalah mengetahui spesies isolat SU-KC1a; menganalisis gen-gen enzim pemecah galaktooligosakarida, laktosa dan galaktosa pada SU-KC1a; serta menganalisis sekuens enzim fosfoketolase pada SU-KC1a. SU-KC1a dari air susu ibu yang diisolasi pada penelitian sebelumnya ditumbuhkan kembali, lalu dikarakterisasi, dan diidentifikasi menggunakan 16S-rRNA sequencing dan Whole Genome Sequencing (WGS). Data 16S-rRNA sequencing diperoleh dengan, esktraksi DNA, PCR, dan dilakukan sequencing, kemudian dianalisis dengan BLAST dan Clustal Omega. Data WGS SU-KC1a didapatkan dari ekstraksi DNA kemudian dilakukan sequencing dengan Oxford Nanopore Ttechnologies. Program seperti Epi2me, Mauve, BLAST, RAST, dan CAZy, digunakan untuk perakitan genom, polishing genom, re-ordering, anotasi dan analisis gen-gen yang mengkodekan enzim pemecah laktosa dan galaktosa sekaligus menganalisis sekuens enzim fosfoketolase. Hasil karakterisasi dan identifikasi molekuler menunjukkan bahwa SU-KC1a identik dengan Lactiplantibacillus plantarum. L. plantarum SU-KC1a dapat memetabolisme galaktooligosakarida, laktosa dan galaktosa karena mengandung gen seperti LacS, LacA, LacL, LacM, LacR galM, galK, galT, dan galE. Kemudian, hasil analisis sekuens pengkode fosfoketolase pada SU-KC1a mirip dengan L.pentosus MD363. Tapi masih diperlukan uji lebih lanjut. / Probiotics can be found in human breast milk, for example, Lactiplantibacillus plantarum (L. plantarum). L. plantarum known for its ability to degrade many kinds of carbohydrates. The purposes of this study are; indicating the species of SU KC1a; analyzing genes that encoded galactooligosaccharides, lactose, and galactose degrading enzymes in L. plantarum; and analyzing sequences that encoded phosphoketolase enzymes in L. plantarum. SU-KC1a from human breast milk that has been isolate in the previous research was cultured, characterized, and identified using 16S-rRNA sequencing and Whole Genome Sequencing (WGS). 16S-rRNA data was obtained from DNA extraction, PCR, and sequencing. Then, the data were analyzed using BLAST and Clustal Omega. WGS data of SU-KC1a was obtained from DNA extraction and sequencing with Oxford Nanopore Technologies. Programs such as Epi2me, Mauve, RAST, and CAZy, were used to assemble, annotate and analyze genes encoding lactose and galactose degrading enzymes. Whole Genome Sequencing analysis indicates that SU-KC1a shows a similar identity with Lactiplantibacillus plantarum. SU-KC1a can metabolize lactose and galactose because the SU-KC1a’s genome contains genes such as LacS, LacA, LacL, LacM, galM, galK, and LacR. Also, the sequence that encodes phosphoketolase enzyme in SU-KC1a is similar to the phosphoketolase enzyme sequence in L.pentosus MD363. But further research is still needed.

Item Type: Thesis (Bachelor)
Creators:
CreatorsNIMEmail
Kindangen, WulanNIM01113170013wkindangen@gmail.com
Contributors:
ContributionContributorsNIDN/NIDKEmail
Thesis advisorJan, Tan TjieNIDN08808640017tjiejan.tan@gmail.com
Thesis advisorVictor, HansNIDN0309049301hvic.uphbiotech@gmail.com
Uncontrolled Keywords: bioinformatika; Whole Genome Sequencing; Lactiplantibacillus plantarum; laktosa dan galaktosa; fosfoketolase
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: University Subject > Current > Faculty/School - UPH Karawaci > Faculty of Science and Technology > Biology
Current > Faculty/School - UPH Karawaci > Faculty of Science and Technology > Biology
Depositing User: Users 4975 not found.
Date Deposited: 24 Sep 2021 03:05
Last Modified: 09 Mar 2022 04:08
URI: http://repository.uph.edu/id/eprint/42542

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