Ketut Ratnayani
Program Studi Kimia FMIPA Universitas Udayana, Bukit Jimbaran, Bali-Indonesia

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PENENTUAN NILAI KM DAN VMAKS DARI ENZIM PAPAIN DALAM HIDROLISIS KONSENTRAT PROTEIN AMPAS TAHU Oka Ratnayani; Sakinatul Fatimah; Ketut Ratnayani; Ni Made Puspawati
CAKRA KIMIA (Indonesian E-Journal of Applied Chemistry) Vol 10 No 2 (2022): Cakra Kimia (Indonesian E-Journal of Applied Chemistry)
Publisher : Graduate Program of Applied Chemistry, Udayana University, Bali-INDONESIA

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Abstract

ABSTRAK: Sisa kandungan protein dalam ampas tahu yang masih cukup tinggi perlu dioptimalkan pemanfaatannya dengan cara mengisolasi komponen protein tersebut sehingga diperoleh konsentrat protein ampas tahu dan digunakan sebagi substrat untuk produksi hidrolisat protein. Penelitian ini bertujuan untuk menentukan kemampuan enzim papain dalam menghidrolisis konsentrat protein ampas tahu yaitu dengan menentukan nilai Km (Konstanta Michaelis-Menten) dan Vmaks (Kecepatan Maksimum). Tahap penelitian diawali isolasi protein total sehingga diperoleh konsentrat proteinnya.yang selanjutnya dihidrolisis secara parsial dengan enzim papain sehingga diperoleh produk hidrolisat protein. Proses isolasi protein ampas tahu menggunakan sampel ampas tahu segar dengan metode ekstraksi alkalik dilanjutkan dengan presipitasi isoelektrik mampu menghasilkan serbuk konsentrat protein dengan nilai kadar protein mencapai 50,36% sehingga memenuhi syarat sebagi konsentrat protein. Hasil penentuan nilai Km dan Vmaks enzim papain dalam menghidrolisis substrat konsentrat protein ampas tahu memperoleh nilai Km sebesar 237,0229 mg/mL dan nilai Vmaks sebesar 5,1975 mg/mL/menit. Hal ini menunjukkan bahwa enzim papain mampu menghidrolisis konsentrat protein ampas tahu sehingga menghasilkan produk hidrolisat protein ampas tahu. ABSTRACT: The remaining protein content in tofu dregs is still quite high, so that its utilization can be optimized by isolating the protein component to obtain the protein concentrate that can be used as a substrate for the production of protein hydrolysates. This study aimed to determine the ability of the papain enzyme to hydrolyze the protein concentrate of tofu dregs through determining the values ??of Km (Michaelis-Menten constant) and Vmax (Maximum Speed). The research steps began with the total protein isolation to obtain the protein concentrate, which was then partially hydrolyzed with papain enzymes to produce protein hydrolysate. The isolation of the tofu dregs protein from fresh tofu dregs samples was done by using an alkaline extraction method followed by isoelectric precipitation resulted in protein concentrate powder with a protein content value of 50.36%, so that it qualified as a protein concentrate. The determination of Km and Vmax values of the papain enzyme in hydrolyzing the protein concentrate substrate of tofu dregs obtained a Km value of 237.0229 mg/mL and a Vmax value of 5.1975 mg/mL/minute. This showed that the papain enzyme was able to hydrolyze the tofu dregs protein concentrate to produce a tofu protein hydrolyzate product.
HIDROLISIS PROTEIN KECAMBAH KACANG MERAH MENGGUNAKAN ENZIM PAPAIN DENGAN VARIASI RASIO ENZIM-SUBSTRAT Oka Ratnayani; Made Ririn Dwi Rahayu; Ketut Ratnayani; Ni Komang Ariati
CAKRA KIMIA (Indonesian E-Journal of Applied Chemistry) Vol 11 No 1 (2023): Cakra Kimia (Indonesian E-Journal of Applied Chemistry)
Publisher : Graduate Program of Applied Chemistry, Udayana University, Bali-INDONESIA

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Abstract

ABSTRAK: Hidrolisis protein secara enzimatik dapat memproduksi hidrolisat protein yang akan menghasilkan peptida rantai pendek dan asam amino bebas, sehingga memiliki nilai gizi dan protein terlarut lebih tinggi dan lebih mudah dicerna. Penelitian akhir-akhir ini juga menunjukkan bahwa kecambah kacang merah (Phaseolus vulgaris L.) memiliki kadar protein terlarut lebih tinggi dibandingkan dengan kacang merah yang belum dikecambahkan. Maka, pada penelitian ini, kecambah kacang merah dijadikan substrat untuk menghasilkan hidrolisat protein menggunakan enzim papain yang relatif mudah didapat, tidak ada reaksi samping, tidak toksik, relatif tahan terhadap suhu, dan memiliki daya katalitik yang tinggi. Selain itu, salah satu faktor yang memengaruhi proses hidrolisis protein adalah rasio enzim-substrat (rasio E/S). Sehingga, tujuan dari penelitian ini adalah untuk mengetahui kadar ?-amino bebas, protein terlarut, dan derajat hidrolisis dari hidrolisat protein kecambah kacang merah hasil hidrolisis menggunakan variasi rasio enzim-substrat. Metode yang digunakan yaitu metode Ninhidrin untuk penentuan kadar ?- amino bebas, metode Biuret untuk penentuan kadar protein terlarut dan metode SN-TCA (Trichloroacetic acid-soluble nitrogen) untuk penentuan derajat hidrolisis. Adapun variasi rasio E/S yang digunakan yaitu 3, 4, 5, dan 6%. Hasilnya menunjukkan bahwa semakin tinggi rasio enzim-substrat, semakin meningkat pula nilai kadar ?-amino bebas, kadar protein terlarut, dan derajat hidrolisis dari hidrolisat protein yang dihasilkan. Hasil terbaik yang didapat, yaitu kadar ?- amino bebas, protein terlarut dan derajat hidrolisis masing masing sebesar 0,5536 mg/mL, 2,0726 mg/mL dan 36,02, diperoleh dari hidrolisis menggunakan rasio E/S 6%. ABSTRACT: The enzymatic protein hydrolysis can produce protein hydrolyzate that will bring out short-chain peptides and free amino acids which contain higher nutritional values and soluble protein content as well as easier to digest. Recently, it has also found that red bean sprouts (Phaseolus vulgaris L.) has higher dissolved protein content than red beans without germination. Thus, in this research, red bean sprouts were used as the substrate to produce protein hydrolysates by using papain enzymes. Papain enzymes are easier to obtain, no side reactions, non-toxic, relatively resistant to temperature, and have high catalytic ability. Moreover, one of the essential factors that affect the protein hydrolysis is the enzyme-substrate ratio (E/S ratio). Therefore, the purpose of this study was to determine the levels of free amino acids, soluble protein, and degree of hydrolysis of the protein hydrolyzate obtained from the hydrolysis of the red bean sprouts using various enzyme-substrate (E/S) ratio. The methods used were Ninhydrin method for determining the free amino acid levels, Biuret test for the soluble protein content and SN-TCA (Trichloroacetic acid-soluble nitrogen) method for the degree of hydrolysis. The variations of the E/S ratio used were 3, 4, 5, and 6%. The results showed that the higher the enzyme-substrate ratio, the higher the value of free amino acids, soluble protein, and degree of hydrolysis in the protein hydrolysate obtained. The highest values of free amino acid, the soluble protein content and the hydrolysis degree of the protein hydrolysate was 0.5536 mg/mL, 2.0726 mg/mL and 36.02%, respectively, obtained by using the hydrolysis with the E/S ratio of 6%.