Palupi, Hapsari Titi and STP, MP, Prof. Dr. Teti Estiasih and DEA, Prof. Dr. Ir. Yunianta and MSc. Ph.D, Ir. Aji Sutrisno (2022) Karakteristik Fisikokimia dan Fungsional Pati dan Konsentrat Protein Kacang Koro Kratok (Phaseolus lunatus L.) dari Pemisahan Simultan Berbasis pH. Doktor thesis, Universitas Brawijaya.
Abstract
Kacang koro kratok (Phaseolus lunatus L.) merupakan bahan pangan dengan profile nutrisi yang baik dan dapat tumbuh di lahan marginal. Kacang jenis ini belum banyak termanfaatkan (underutilized) dan masih sedikit dilakukan penelitian tentang potensinya. Sebagai sumber protein dapat digunakan sebagai bahan baku pembuatan konsentrat protein. Saat ini terdapat perhatian yang cukup besar, tidak hanya mendapatkan konsentrat protein tetapi juga pati. Ekstraksi basa-pengendapan isoelektrik adalah metode ekstraksi yang digunakan untuk memisahkan pati dan protein terlarut, selanjutnya digunakan untuk mendapatkan protein. pH merupakan faktor penting pada proses ekstraksi pati dan protein, karena dapat mempengaruhi komposisi kimia, struktur dan sifat fungsional dari pati dan protein yang dihasilkan. Pati dan konsentrat protein dapat digunakan sebagai bahan baku pengolahan pangan. Tujuan penelitian adalah mengembangkan teknologi pengolahan untuk pemanfaatan kacang koro kratok, khususnya komponen pati dan protein, dan mempelajari lebih lanjut terkait sifat fisikokimia dan fungsional dari pati dan protein yang dipisahkan. Penelitian ini terdiri atas 3 tahap. Penelitian tahap 1: Karakterisasi biji dan tepung kacang koro kratok. Karakterisasi biji kacang koro kratok meliputi: komponen gizi, anti gizi dan toksin HCN, asam amino, pengukuran fraksi protein berdasarkan kelarutan, serta profil protein dengan elektroforesis SDS-PAGE. Karakterisasi tepung meliputi: komponen gizi, anti gizi dan toksin HCN, sifat pemastaan dan fungsional. Masing-masing parameter diulang 3 kali. Penelitian Tahap 2: Pemisahan pati kacang koro kratok dan protein terlarut dengan pH ekstraksi basa. Rancangan penelitian yang digunakan adalah Rancangan Acak Kelompok (RAK) dengan perlakuan pH ekstraksi basa yaitu: pH 7, 8, 9, 10, dan 11 serta masing-masing diulang 3 kali. Respon yang diamati di pati yaitu: proksimat, rendemen, kadar pati, amilosa, warna, derajat putih, sifat pemastaan, kelarutan, daya pembengkakan, serta daya serap air dan minyak, morfologi granula, pola difraksi sinar X, dan gugus fungsi. Respon yang diamati di protein terlarut yaitu: kadar protein terlarut dan fraksi protein. Penelitian Tahap 3: Pembuatan konsentrat protein kacang koro kratok berdasarkan pH pengendapan asam. Rancangan penelitian yang digunakan Rancangan Acak Kelompok (RAK) dengan perlakuan pH pengendapan asam yaitu: pH 3,5; 4; 4,5; 5; dan 5,5; serta pengulangan 3 kali. Respon diamati di konsentrat protein yaitu: senyawa proksimat, rendemen, warna, asam amino, mikrostruktur, profil elektroforesis protein-gel doc, sifat termal, kelarutan, WHC, OHC, kapasitas dan stabilitas buih, serta EAI dan ESI. Hasil penelitian tahap 1 menunjukkan kacang koro kratok merupakan sumber nutrisi baik yaitu mengandung protein 15,93%, rendah lemak (1,13%), dan relatif tinggi karbohidrat kompleks (68,02%) yang terdiri dari pati dan serat pangan. Kacang koro kratok kaya akan asam amino esensial seperti lisin, leusin dan fenilalanin. Fraksi protein berdasarkan kelarutan di kacang koro kratok adalah globulin (56,20%) diikuti glutelin (22,69%) dan albumin (18,47%), sedangkan prolamin ditemukan dalam jumlah yang sangat rendah (3,14%). Fraksi globulin utama adalah vicilin dengan BM 15, 24, 35, dan 48 kDa. Kacang koro kratok juga mengandung komponen anti gizi dan toksin. Pengolahan menjadi tepung meningkatkan kadar protein (17,77%), pati (46,34%), dan menurunkan komponen serat (13,34%). Senyawa anti gizi dan toksin juga menurun. Tepung kacang koro kratok memiliki viskositas puncak, suhu pemastaan tinggi dan viskositas breakdown rendah, menunjukkan tepung memiliki stabilitas pasta yang baik dan ketahanan geser yang kuat. Hasil penelitian tahap 2 menunjukkan pH ekstraksi berpengaruh nyata terhadap sifat fisikokimia, pemastaan, morfologi, dan derajat kristalinitas. Semakin tinggi pH ekstraksi basa maka kemurnian pati kacang koro kratok semakin meningkat. Pati kacang koro kratok memiliki kadar amilosa yaitu 44,03-50,65% dan kadar amilosa berpengaruh pada daya pembengkakan, kelarutan, daya serap air dan minyak, sifat pemastaan dan derajat kristalinitas. Karakteristik morfologi pati menunjukkan sedikit kerusakan korosif di permukaan granula pada pH ekstraksi 11. Perlakuan pH basa tidak menyebabkan perubahan pola difraksi sinar X dan menunjukkan kristal pati tipe CA. Gugus fungsional memperlihatkan tren spektrum sama untuk semua pati. Secara umum pati memiliki kadar amilosa, viskositas puncak dan temperatur gelatinisasi tinggi serta viskositas setback rendah menunjukkan pati kacang koro kratok memiliki daya tahan tinggi terhadap pembengkakan, stabilitas tinggi terhadap panas dan gaya mekanis, serta sumber pati resisten. Dari karakteristik diatas, maka pati kacang koro kratok berguna untuk bahan baku untuk sejumlah aplikasi makanan. Untuk kadar protein terlarut, semakin tinggi pH ekstraksi basa maka terjadi peningkatan signifikan terhadap kadar protein terlarut di filtrat, dan diperoleh kadar protein terlarut tertinggi (17,91 mg/ml) di pH ekstraksi 11 pH ekstraksi basa berpengaruh terhadap komposisi fraksi protein. Filtrat dari pH ekstraksi 11 digunakan untuk tahap 3. Hasil penelitian tahap 3 menunjukkan pengaturan pH ekstraksi basa dan pengendapan asam dapat menjadi strategi yang efektif untuk mengisolasi protein kacang koro kratok. pH pengendapan asam berpengaruh nyata terhadap kadar protein, rendemen, dan sifat fungsional konsentrat protein kacang koro kratok. pH pengendapan 4,5 menghasilkan rendemen dan kadar protein tertinggi. Perlakuan pH pengendapan asam menghasilkan konsentrat protein yang memiliki variasi pada komposisi asam amino. Semua konsentrat protein kacang koro kratok menunjukkan struktur kristal melalui pengamatan SEM dan memiliki stabilitas termal tinggi. Vicilin merupakan fraksi protein dominan di semua konsentrat protein kacang koro kratok dengan BM 26, 29, 35, 45, 57, dan 72 kDa, dan memiliki persentase berbeda untuk masing-masing perlakuan. Variasi komposisi asam amino dan konformasi protein berpengaruh terhadap terhadap sifat fungsional konsentrat protein. pH pengendapan asam 3,5 memiliki kelarutan dan WHC lebih tinggi, sedangkan pH pengendapan asam 4,5 memiliki OHC, kapasitas buih, EAI dan ESI yang lebih tinggi. Berdasarkan karakteristik fungsional konsentrat protein kacang koro kratok dapat dimanfaatkan sebagai bahan baku produk makanan. Untuk penggunaannya sebagai bahan baku di pengolahan makanan maka diperlukan penelitian lebih lanjut terkait aplikasi dari pati kacang koro kratok pada pembuatan produk yaitu ekstrudat, makanan kaleng, sosis, saus dan mie. Sedangkan konsentrat protein kacang koro kratok dapat dilakukan penelitian terkait pemanfaatannya sebagai bahan substitusi untuk produk sosis dan bakery serta menganalisis kualitas produk yang dihasilkan.
English Abstract
Lima bean or Koro kratok (Phaseolus lunatus L.) is a food ingredient with a good nutritional profile and grows in marginal land and environment. Lima bean is still underutilized and its potential is under-researched. As a protein source, it can be used as an ingredient for making protein concentrates. Currently there is sufficient attention, not only to having the protein concentrates, but also to having starch sources. In the food industry, alkaline extraction isoelectric precipitating (AE-IEP) technique is commonly applied approaches to separate starch and soluble protein, which is then used to obtain protein. pH is an important factor in the starch and protein extraction process because it alters protein structure and, subsequently, its function. Starch and protein concentrates can be used as raw materials for food processing. The aim of the research is to develop technological process for the for complete use of lima bean, specifically its protein and starch fractions, and effects of extraction pH and acid precipitation pH on the physicochemical and functional properties of starch and protein separated simultaneously. This research consists of 3 steps. Step 1: Characterization of seeds and flour of Lima bean. Characterization of Lima bean included nutritional composition, anti-nutritional and toxin components, amino acids, protein fraction, and protein profile by electrophoresis SDS-PAGE. The characterization of flour: nutritional, anti-nutritional and toxin components, pasting and functional properties. Each parameter was repeated 3 times. Step 2: Separation of lima bean starch and soluble protein by alkaline extraction pH. Using a Randomized Block Design (RAK), independent factors are extraction pH at pH 7, 8, 9, 10, and 11, each repeated 3 times. The responses observed in starch are proximate, yield, starch content, amylose, colour, whiteness, pasting properties, solubility, swelling power, water and oil absorption, granule morphology, X-ray diffraction pattern, and FTIR. Responses observed in soluble protein are soluble protein content and protein fraction. Step 3: Process of acid precipitation at different pH to obtained protein concentrate. Using a Randomized Block Design (RAK), independent factors are acid precipitation pH at 3.5; 4; 4.5; 5; and 5.5. Repeated 3 times. Responses observed are: proximate compounds, yield, color, amino acids, microstructure, electrophoretic profile of protein-gel doc, thermal properties, solubility, EAI and ESI, WHC, OHC, foam capacity and stability. The results of step 1 showed that lima bean is a good source of nutrition, having a moderate concentration of protein (15.93%), low concentration of fat (1.13%), and relatively high in complex carbohydrates (68.02%) consisting of starch and dietary fibre. Lima bean is also rich in essential amino acids such as lysine, leucine and phenylalanine. The storage protein fraction in Koro kratok was globulin (56.20%) followed by glutelin (22.69%) and albumin (18.47%), while prolamins were found in very low amounts (3.14%). The major globulin fraction was vicilin with MW of 48, 35, 24, 15 kDa. Several anti-nutritional compounds were found: trypsin, phytic acid, saponins, TPC, oligosaccharides, and toxic compounds as HCN. Processing is needed to reduce anti-nutritional components and increase their utilization. Processing of seed into flour increased the protein content (17.77%), starch (46.34%) and decreased the dietary fibre (13.34%). Anti-nutritional compounds and HCN decrease. Lima bean flour has peak viscosity, high pasting temperature and low breakdown viscosity, indicating the flour has good paste stability and shear resistance. The results of step 2 The present research has provided an overview into the impact of alkali extraction. pH on various properties of lima bean starches, that extraction pH has significant effects on the physicochemical, morphological, crystallinity, pasting, and functional characteristics of lima bean starches. Increasing extraction pH showed a greater level of purity. The starch of lima bean has higher amylose content of 44.03 to 50.65%. The amylose content was observed to have a slight decrease in higher extraction pH. Amylose content affected the morphological, crystallinity, pasting, and functional properties of starch. Morphological characteristics of alkaline treated starches showed deformation with the corrosive damage of granular surface at higher extraction pH. Extraction pH caused no changes in the XRD pattern but caused a slight change in intensity and crystallinity. The pasting properties of the lima bean starches changed significantly with different extraction pH. However, in general, starch has a high pasting temperature and lower values of breakdown viscosity. High swelling power, WHC, OAC, and pasting properties are all excellent that make it provides the opportunity to utilize the potential of starches in unexplored for industrial applications. The higher the extraction pH, there was a significant increase in the dissolved protein content in the filtrate, and the highest dissolved protein content (17.91 mg/ml) was obtained at the extraction pH 11. Different extraction pH affected the composition of protein fractions. The results of step 3 showed that pH adjustment of alkaline extraction and acid precipitation could be an effective strategy for isolating lima bean protein. The acid precipitation pH had a significant effect on the protein content, yield, and functional properties of lima bean’s protein concentrate. Acid precipitation pH of 4.5 resulted in the highest yield and protein content. Acid precipitation pH produces protein concentrates that have variations in amino acid composition. All lima bean protein concentrates showed a compact crystal structure by SEM observation and had high the MW of 72, 57, 45, 35, 29 and 26 kDa, but showed different levels. Variations in amino acid composition and protein conformation affect the functional properties of protein concentrates. Precipitation pH of 3.5 has higher solubility and WHC, while pH 4.5 has higher OHC, foam capacity, EAI and ESI. Based on the functional characteristics of lima bean protein concentrates, it can be used as raw material for the development of new food products and the functional characteristics affect its use. However, further research is considered important to expand the application of these ingredients in food formulations. it is necessary to conduct further research related to the application of the use of lima bean starch in food products, ex: extrudates, canned food, sausages, sauces and noodles, as well as the utilization of protein concentrate of lima beans as a substitute for sausage and bakery products and analyse the characteristics of the resulting product.
| Item Type: | Thesis (Doktor) |
|---|---|
| Identification Number: | 0623100001 |
| Uncontrolled Keywords: | ekstraksi basa-pengendapan asam, kacang koro kratok, konsentrat protein, pati, pH., alkaline extraction-acid precipitation, lima bean, pH, protein concentrate, starch. |
| Subjects: | 300 Social sciences > 338 Production > 338.1 Agriculture > 338.16 Production efficiency |
| Divisions: | S2/S3 > Doktor Teknologi Industri Pertanian, Fakultas Teknologi Pertanian |
| Depositing User: | soegeng sugeng |
| Date Deposited: | 17 May 2023 03:10 |
| Last Modified: | 19 May 2023 01:34 |
| URI: | http://repository.ub.ac.id/id/eprint/199774 |
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