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All Journal CAKRA KIMIA (Indonesian E-Journal of Applied Chemistry) Jurnal Kimia (Journal of Chemistry) International Journal of Biosciences and Biotechnology INDONESIAN JOURNAL OF BIOMEDICAL SCIENCES The Journal of Pure and Applied Chemistry Research Indonesian Journal of Chemistry Journal of Health Sciences and Medicine

Ketut Ratnayani

Program Studi Kimia, Fakultas MIPA Universitas Udayana, Bali-Indonesia

Author-ID : 2921952

Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Electrical & Electronics Engineering Environmental Science Materials Science & Nanotechnology Medicine & Pharmacology

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DESAIN PRIMER UNTUK AMPLIFIKASI GEN katG MULTIDRUG RESISTANCE TUBERCULOSIS (MDR-TB) DENGAN METODE POLYMERASE CHAIN REACTION (PCR) Putu Tedi Suryadi; Ketut Ratnayani; Sagung Chandra Yowani
Jurnal Kimia (Journal of Chemistry) Vol. 8, No. 1 Januari 2014
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

Tuberculosis, the world’s major diseases, is one of the emerging infectious disease. The tuberculosis problem has become complicated and burdensome due to the emergence of drug resistant such as isoniazid (INH) resistant strains of Mycobacterium tuberculosis. Mutation in katG gene is the main mechanism of INH-resistance in most strains. Amplification of M. tuberculosis katG gene was performance by using PCR for detect the mutation. A pair of specific PCR primers (forward and reverse) was the most important factor to limit the target region of amplification. Primer designing is preceedly carried out for producing the specific primer desired. The aim of this study was to design the specific primers for a fragment of katG gene. In silico primer design was carried out by using Clone Manager Suite 6. DNA sequence template used in this primer design was downloaded from www.ncbi.nml.nih.gov. M. tuberculosis H37Rv katG gene with genbank code X68081.1 was choosen. This study was successfully designed the forward primer (5' GAAGTACGGCAAGAAGCTCTC 3') and reverse one (5' CGTGATCCGCTCATAGATCG 3') which was length of 21 and 20 nucleotide, respectively. These pair of primers were meet the requirement of a good primer include primer length, Tm value, percentage of GC content, no hairpins, limited dimers and runs. In conclusion, the result of this research showed that the primer designed were acceptable to amplify the fragment of 724 bp of katG gene in silico.

KAJIAN PENGARUH VARIASI KONSENTRASI ASAM SITRAT TERHADAP KEKUATAN GEL PRODUK GELATIN KULIT AYAM BROILER DIKAITKAN DENGAN POLA PROTEINNYA Tutut Hardikawati; Ni Made Puspawati; Ketut Ratnayani
Jurnal Kimia (Journal of Chemistry) Vol. 10, No. 1 Januari 2016
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

Gelatin is a biopolymer that can be generated from partially hydrolysis of collagen tissue. Extraction of gelatin consists of pretreatment and thermal extraction steps. Pretreatment process used sodium hydroxide to remove non collagen protein in matrix sample, sulfuric acid to demineralize, and citric acid to hydrolyse. The aim of this research was to study the effect of variation in concentrations of citric acid used in hydrolysis process on the gel strength and protein profile of gelatin products extracted from broiler chicken skin. The variation of the concentration citric acid used was 0.7 % (GA); 1.5% (GB); and 3.0% b/v (GC) respectively. The gel strength was measured using CT3 Texture Analyzer and protein profile of gelatin product was analyzed by SDS-PAGE method. The result showed that variation in concentration of citric acid used in the pretreatment process affected the gel strength and protein profile of gelatin product. Increasing the concentration of citric acid used in pretreatment process decreased the gel strength and molecular weight of gelatin product. Gel strength of each gelatin product was 265.81 g bloom for GA ; 196.05 g bloom for GB (1.5%), and 35.32 g bloom for GC (3.0 %) respectively. The electropherogram of both GA (0.7%) and GB (1.5%) revealed similar pattern of protein bands but the thickness of each bands was different. On the other hands, GC (3.0%) did not show any protein bands on the eletropherogram. The best gelatin product obtained in this experiment was found by using 0.7 % b/v citric acid (GA) in the pretreatment process. The gelatin product (GA) had characteristics as follows: yield 15.73%; moisture 7.30%; ash 0.51%; protein content 97.95%; fat content 0.62%; gel strength 265. 81 g bloom and thicker protein bands than others.

ISOLASI DAN IDENTIFIKASI SENYAWA GOLONGAN FLAVONOID DARI MADU K ELENGKENG (Nephelium longata L.) Ida Ayu Raka Astiti Asih; Ketut Ratnayani; Ida Bagus Swardana
Jurnal Kimia (Journal of Chemistry) Vol. 6, No. 1 Januari 2012
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (203.213 KB)

The determination of anti free radical activity on longan honey (Nephelium longata L.) by DPPH method using UV-Vis sphectrophotometry and identification of chemical compound in non polar and semi polar fraction have been done. Longan honey was diluted with methanol and then partied by n-hexane and ethyl acetate. The absorbance was measured at 497 nm, 517 nm, and 537 nm for the DPPH concentration of : 0,001%, 0,002%, 0,003%, and 0,004% and the chemical compound was identified by phytochemical method.The result showed that part of n-hexane and ethyl acetate probably consist of chemical compound of isoflavone and value of anti free radical activity on longan honey in semi polar fraction was higher than in non polar fraction which were 91,71% and 77,68% at DPPH concentration of 0,001% (b/v).

KAPASITAS ADSORPSI BEBERAPA JENIS KULIT PISANG TERAKTIVASI NaOH SEBAGAI ADSORBEN LOGAM TIMBAL (Pb) Putu Eka Purnama; I Gusti Ayu Kunti Sri Panca Dewi; Ketut Ratnayani
Jurnal Kimia (Journal of Chemistry) Vol. 9, No. 2 Juli 2015
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

Adsorption capacity of various types of banana skin (green, kepok and susu) activated with NaOH as adsorbents for lead (Pb) has been studied. This research consisted of several steps included determination of the surface area of activated and unactivated adsorbents, equilibrium time, adsorption isotherms and adsorption capacity of adsorbents from green, kepok and susu banana skins activated with NaOH . The results showed that adsorbents from green, kepok and susu banana skin activated with NaOH had surface were of 36.2181 m2/g, 35.5531 m2/g and 35.8378 m2/g respectively. On the other hand, the surface area of unactivated adsorbents of green, kepok and susu banana skin were 35.3105 m2/g, 35.3199 m2/g, and 35.7928 m2/g respectively. Equilibrium time for green, kepok and susu banana skin adsorbents activated with NaOH were 30; 30 and 90 minutes. Adsorption isotherms of adsorbents from green, kepok and susu banana skin activated with NaOH were at concentration 100 ppm. Adsorption capacity of activated adsorbents from green banana, kepok banana and susu banana skin were 7.022 mg/g, 5.3078 mg/g and 6.6850 mg/g respectively.

PENGARUH PENAMBAHAN SUSU SKIM TERHADAP HASIL DNA METAGENOMIK DIISOLASI DARI TANAH HUTAN MANGROVE Ni Putu Frida Oktaningtias Widiarthi; Ketut Ratnayani; I Nengah Wirajana
Jurnal Kimia (Journal of Chemistry) Vol. 8, No. 1 Januari 2014
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

Cell lysis is the most important step for quality and quantity of metagenomic DNA isolated from an environmental samples. The aim of the research was to compare the quality (integrity and purity) of the metagenomic DNA isolated using the direct cell lysis method from mangrove forest soil with and without skim milk. The total of metagenomic DNA isolated from mangrove forest soil result was analyzed by the spectrophotometric UV-Vis method at ? 230, 260, and 280 nm; and also by using the agarose gel electrophoresis. The results showed that metagenomic DNA can be isolated from mangrove forest soil. The agarose gel electrophoresis results showed that the total DNA quality obtained by the direct cell lysis using buffer lysis with skim milk was relatively less fragmented and the band intensity of DNA was higher compared with direct cell lysis using buffer lysis without skim milk. The results of spectrophotometry indicated that the purity of DNA isolated with and without skim milk was not significantly different against the humic acid (ratio on A260/230). As shown by the A260/280 ratio, the total DNA isolated without skim milk had higher purity level than with skim milk.

ISOLATION OF PROTEASE ENZYME FROM CHAYOTE FRUIT (Sechium edule (Jacq.) Sw.) WITH AMMONIUM SULFATE FRACTINATION METHOD Ketut Ratnayani; Lia Kusumaningrum
International Journal of Biosciences and Biotechnology Vol 2 No 2 (2015)
Publisher : Central Laboratory for Genetic Resource and Molecular Biology, Faculty of Agriculture, Udayana University in cooperation with Asia-Oceania Bioscience and Biotechnology Consortium (AOBBC)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (409.676 KB)

Protease is an enzyme that is capable to hydrolyze (breakdown) protein molecules intosimpler compounds such as small peptides and amino acids. The aim of the research was toisolate protease enzyme from chayote (Sechium edule (Jacq.) Sw.) using fractinationammonium sulfate method and to find out the optimum saturation level of the ammoniumsulfate. P rotease activity examination of each fraction of ammonium sulfate was performedusing Anson method. Protein content assay was determined using Biuret method. The resultsshowed that crude extract protease of chayote had specific activity of 3,7338 x 10-3 U/mg. Theoptimal saturation levels of ammonium sulfate for protease chayote precipitation was 40-50%. At this saturation level, the highest enzyme spesific activity were 16,00 x 10-3 U/mg,with four times purifying of protease enzyme from the crude extract protease.

SCREENING POTENTIAL ANTIOXIDANT AND ANTIBACTERIAL ACTIVITIES OF PROTEIN HYDROLYSATES DERIVED FROM GERMINATED LABLAB BEAN, PIGEON PEA AND KIDNEY BEAN Ketut Ratnayani; Indriani Wisnu Susanto Panjaitan; Ni Made Puspawati
Journal of Health Sciences and Medicine Vol 1 No 1 (2017): JHSM (Febuary 2017)
Publisher : Institute for Research and Community Services Udayana University

Abstract Protein hydrolysate contains a mixture of various lengths of short peptides chain and free amino acids that may excert biological activities. This research aims to screen potential antioxidant and antibacterial activities of protein hydrolysate produced from three kinds of germinated beans i.e. lablab bean (Lablab purpureus), pigeon pea (Cajanus cajan (L.) Millsp) and kidney bean (Phaseolus vulgaris) through enzymatic hydrolysis process. The steps of research included germination process of the beans prior to total protein isolation, enzymatic hydrolysis of total protein isolates using pancreatin enzyme, evaluation of in vitro antioxidant activity of the hydrolysates protein using DPPH (1,1-diphenyl-2-picryl hydrazyl) method, and antibaterial activity testing towards Eschericia coli and Staphyllococcus aureus bacteria. The results revealed that pancreatine enzyme was able to hydrolyse germinated protein of lablab bean, pigeon pea and kidney bean at the experiment condition applied with degree of hydrolysis 34.12%, 27.44%, and 30,93% respectively. It was also found that protein hydrolysates of lablab bean, pigeon pea, and kidney bean demonstrated antioxidant activity which percentage radical DPPH scavenging activity of 84.02%, 68.97% and 67.89 %. On the other hand, all of those protein hydrolysates did not show any antibacterial activity towards Eschericia coli and Staphyllococcus aureus bacteria.

Angiotensin Converting Enzyme (ACE) Inhibitory Activity of Peptide Fraction of Germinated Pigeon Pea (Cajanus cajan (L.) Millsp.) Ketut Ratnayani; I Ketut Suter; Nyoman Semadi Antara; I Nengah Kencana Putra
Indonesian Journal of Chemistry Vol 19, No 4 (2019)
Publisher : Universitas Gadjah Mada

During the germination process, seeds can release various types of peptides due to the degradation of storage proteins. Some of these peptides can have biological activity (bioactive peptides). The objective of this study was to determine the ACE inhibitory activity of germinated pigeon pea peptide extract at various germination times and to carry out the fractionation to the extract to get the most active peptide fraction. The results showed that the highest activity of peptide extract was found on the 4th-day germination of pigeon pea with an IC50 value of 63.46 μg/mL. The peptide extract was further fractionated by centrifugal ultrafiltration method and it was found that the peptide fraction < 3 kDa had the highest ACE inhibitory activity with an IC50 value of 57.79 μg/mL. The result of identification with the LCMS method to the fraction was able to detect 4 types of the peptide with a molecular weight of 230.304, 294.303, 441.436, and 570.591 Da. These results suggested that the peptide fraction of germinated pigeon pea has the potency as an ACE inhibitory nutraceutical.

Effect of Solvent Type and Germination Time on The Level of Free Amino Acid and Peptides of Germinated Pigeon Pea (Cajanus cajan (L.) Millsp) Extract Ketut Ratnayani; Ni Wayan Wisaniyasa; Ni Putu Wiwik Oktayuni
The Journal of Pure and Applied Chemistry Research Vol 11, No 2 (2022): Edition May-August 2022
Publisher : Chemistry Department, The University of Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jpacr.2022.011.02.672

Germination can activate the degradation of storage protein in legumes releasing peptides and free amino acids for seed growth. These compounds have many benefits in many fields, especially in food and health. This study aimed to determine the effect of solvent type and germination time on the level of free amino acids (FAA) and the level of peptides (MW < 10 kDa) of germinated pigeon pea extract. The extraction of free amino acids and the dissolved protein from germinated pigeon pea flour was carried out using two kinds of solvents, namely water, and 0.1 N HCl. The variation of germination times of pigeon pea seeds was carried out at 12, 24, 36, 48, 60, 72, 84, and 96 hours. The level of FAA was determined spectrophotometrically using the ninhydrin method after the deproteination step, while the level of peptides was determined spectrophotometrically using the biuret method. The results showed that using 0.1 N HCl as a solvent produced a higher level of FAA and peptides in the extract than using water. The increase of germination time can increase the level of FAA with the peak FAA level achieved on 36 hours. The increase of germination time can increase the level of peptides with the peak of peptides level achieved on 84 hours of germination times. These results showed that seed germination is a potential method for producing free amino acids and peptides which accumulate at a specific time.

MONITORING HIDROLISIS PROTEIN KECAMBAH KACANG TUNGGAK (Vigna unguiculata L.) OLEH ENZIM ALKALASE PADA VARIASI WAKTU DAN RASIO ENZIM-SUBSTRAT Ratnayani, K.; Listiyanti, N. K. L.; Ariati, N. K.; Laksmiwati, A. A. I. A. M.
Jurnal Kimia (Journal of Chemistry) Vol. 18, No.2, Juli 2024
Publisher : Program Studi Kimia, FMIPA, Universitas Udayana (Program of Study in Chemistry, Faculty of Mathematics and Natural Sciences, Udayana University), Bali, Indonesia

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Kacang tunggak dengan kandungan protein 18,3-35% berpotensi digunakan sebagai substrat protein dalam pembuatan hidrolisat protein nabati. Kecambah kacang tunggak cukup mudah diperoleh di pasaran (khususnya di wilayah Bali) yang lebih dikenal sebagai kacang mentik. Penelitian ini bertujuan melakukan monitoring berjalannya reaksi hidrolisis konsentrat protein kecambah kacang tunggak yang dikatalisis oleh enzim alkalase, dengan perlakuan variasi waktu hidrolisis dan variasi rasio enzim-substrat (rasio E/S). Penelitian diawali dengan tahap ekstraksi protein dari tepung kecambah kacang tunggak sehingga diperoleh konsentrat protein. Selanjutnya dilakukan proses hidrolisis terhadap substrat konsentrat protein dengan enzim alkalase menggunakan variasi waktu hidrolisis 0, 1, 2 dan 3 jam serta variasi rasio (E/S) yaitu 0,1%, 1,0%, 2,0% dan 3,0%. Masing-masing hidrolisat protein yang diperoleh dimonitor keberhasilannya dalam hidrolisis berdasarkan parameter kadar kadar ?-amino bebas, nilai derajat hidrolisis dan kadar protein terlarutnya. Hasil ekstraksi protein mampu menghasilkan konsentrat protein kacang tunggak dengan kadar protein total mencapai 68,11%. Hasil monitoring terhadap perlakuan hidrolisis menunjukkan bahwa peningkatan rasio E/S dan lama waktu hidrolisis mampu meningkatkan kadar ?-amino bebas, kadar protein terlarut, dan nilai derajat hidrolisis (DH%) dari produk hidrolisat protein kecambah kacang tunggak (pada batas variasi perlakuan yang diberikan). Kadar ?-amino bebas tertinggi sebesar 3,9573 mg/mL, kadar protein terlarut tertinggi sebesar 20,9972 mg/mL, dan nilai DH% tertinggi sebesar 93,80% diperoleh menggunakan perlakuan rasio E/S 3% dan waktu hidrolisis selama 3 jam. Kata kunci: alkalase, hidrolisis, kacang tunggak, kecambah, protein ABSTRACT Cowpea usually contains 18.3-35% protein and is potentially used as a protein substrate for preparing vegetable protein hydrolysates. Cowpea sprouts, available in the market (especially in Bali), are known as “kacang mentik”. This study aimed to monitor the progress of the hydrolysis of the cowpea sprout protein concentrate, catalyzed by alcalase enzyme, with the variations of hydrolysis time and enzyme-substrate ratio (E/S ratio). The research began with the protein extraction stage from cowpea sprout flour to obtain the protein concentrate. Furthermore, the hydrolysis was carried out on the protein concentrate substrate with alcalase enzyme using the variations of hydrolysis time of 0, 1, 2, and 3 hours and the E/S ratio of 0.1, 1.0, 2.0, and 3.0%. Each protein hydrolysate obtained was monitored for its success in hydrolysis based on the parameters of soluble protein content, free ?-amino content, and the degree of hydrolysis. The result showed that the protein extraction produced cowpea protein concentrate with a total protein content reaching 68.11%. The monitoring results of the hydrolysis treatment showed that increasing the E/S ratio and the length of hydrolysis time were able to increase the free ?-amino content, soluble protein content, and the degree of hydrolysis (DH%) value of the cowpea sprout protein hydrolysate product (at the limit of the treatment variations given). The highest value of the free ?-amino content of 3.9573 mg/mL, the soluble protein content of 20.9972 mg/mL, and the DH of 93.80% was obtained when using the 3% E/S ratio treatment and 3 hours of hydrolysis time. Keywords: alcalase, cowpea, hydrolysis, protein, sprout

Co-Authors A. A. Ayu Septri Juwarni A. W. Dwiputri Acintya, N. M. T. C. Amanda Awalia Ramadhani Ana Listya Anak Agung Istri Agung Mayun Laksmiwati Ariati, N. K. Darma Asih Yuliana Deniariasih, N.W. Dewi, I G. A. K. S. P. Dita Rizkiyanti Helen Helda Prastika I G. A. M. A. S. Gitadewi I Gusti Ayu Kunti Sri Panca Dewi I Ketut Suter I Made Oka Adi Parwata I Nengah Kencana Putra I Nengah Wirajana I Wayan Sudiarta I Wayan Suirta I. A. Preharsini Kusuma I. A. R. Astiti Asih Ida Bagus Swardana Indriani Wisnu Susanto Panjaitan James Sibarani Lia Kusumaningrum Liangky Syane S Listiyanti, N. K. L. M. A. Pratiwi M. Nazib Made Arsa Maman Sudiarto Ni G. A. M. Dwi Adhi Suastuti Ni Komang Ariati Ni Luh Putu Mega Wahyuni Ni Made Puspawati Ni Made Suaniti Ni P. Indah Septian P. Ni Putu Frida Oktaningtias Widiarthi Ni Putu Wiwik Oktayuni Ni Wayan Agustini Ni Wayan Wisaniyasa NYOMAN SEMADI ANTARA Putra, A. A. B. Putu Eka Purnama Putu Lita Astriani Putu Setya Sri Antary Ro’yal Aini Sagun Chandra Yowani Suarsa, I W. Syamsul Arifin Tutut Hardikawati

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