• International Journal of Technology (IJTech)
  • Vol 10, No 8 (2019)

The Roles of Candida tropicalis Toward Peptide and Amino Acid Changes in Cheese Whey Fermentation

Isfari Dinika, Bambang Nurhadi, Nanang Masruchin, Gemilang Lara Utama, Roostita L. Balia

Corresponding email: g.l.utama@unpad.ac.id


Cite this article as:
Dinika, I., Nurhadi, B., Masruchin, N., Balia, R.L., Utama, G.L., 2019. The Roles of Candida tropicalis Toward Peptide and Amino Acid Changes in Cheese Whey Fermentation . International Journal of Technology. Volume 10(8), pp. 1533-1540

237
Downloads
Isfari Dinika Food Technology, Faculty of Agricultural Industrial Technology, Universitas Padjadjaran, Bandung-Sumedang Highway Km. 21, Sumedang 45363, Indonesia
Bambang Nurhadi Food Technology, Faculty of Agricultural Industrial Technology, Universitas Padjadjaran, Bandung-Sumedang Highway Km. 21, Sumedang 45363, Indonesia
Nanang Masruchin Research Center for Biomaterial LIPI, Indonesian Institute of Science. Jakarta-Bogor Highway Km. 47, Bogor 16911, Indonesia
Gemilang Lara Utama 1. Food Technology, Faculty of Agricultural Industrial Technology, Universitas Padjadjaran, Bandung-Sumedang Highway Km. 21, Sumedang 45363, Indonesia 2. Center for Environment and Sustainability Sc
Roostita L. Balia Animal Product Technology, Faculty of Animal Husbandry, Universitas Padjadjaran, Bandung-Sumedang Highway Km. 21, Sumedang 45363, Indonesia
Email to Corresponding Author

Abstract
image

Whey is a by-product of cheese processing and is comprised of nearly 90% of the milk used. The protein content in cheese whey has the potential to create peptide and amino acids which have a functional effect in biological activity. Peptides and amino acids can be produced through fermentation with Candida tropicalis into native whey from cheese whey. The study aims to determine fermentation time in producing peptide and amino acid profiling in the fermentation of native cheese whey by Candida tropicalis. Cheese whey fermented with C. tropicalis was compared to a naturally fermented cheese whey as control at an ambient temperature for 48 hours. Peptide content identified by Folin–Ciocalteu methods and the amino acid profile is determined by high performance liquid chromatography (HPLC). Fermentation results showed that the maximum content of peptides needs a 24-hour fermentation in 10.42 ppm. Peptide content decreased with further fermentation caused by the degradation of peptides into amino acids. The amino acids that increased were aspartate, glutamate, threonine, valine, isoleucine, and lysine, while those that decreased were serine, histidine, glycine, arginine, alanine, tyrosine, and methionine.

Amino acid; Candida tropicalis; Native cheese whey; Peptide content

Introduction

As a by-product of cheese processing, whey can be harmful to the environment because of the value of Biochemical Oxygen Demand (BOD) that can exceed 35,000 ppm and a Chemical Oxygen Demand (COD) of more than 60,000 ppm. Based on that statement, the environmental harm of 4,000 L of whey can be compared to 1,900 L of human feces (Smithers, 2008). In mozzarella cheese production, almost 90% of raw materials become whey. As a result, 100 L of milk can produce 80–90 L of whey (Božani? et al., 2014). Based on the amount and the harmful effect to the environment, whey as an agro-waste biomass needs to be handled before disposal (Hawashi et al., 2019).

Despite being a by-product, whey still has many nutritional benefits. Whey has 55% of the total nutrients contained in milk (Andrade et al., 2017). Specifically, whey contains 93.7% (w/w) water, 0.1–0.5% (w/w) fat, 0.8% (w/w) protein,  4.9% (w/w) lactose,  0.5–0.8% (w/w) ash,  and 0.1–0.4% (w/w) lactic acid (Božani? et al., 2014). Because of those nutritional characteristics, 70% of whey can be utilized to become another product, such as whey powder, that can be used in various products such as pastry, sweets, jams, and melted cheese. The added cost and technology to turn whey into whey powder can be a reason it isn’t often applied in rural areas. That’s why only 30% of whey is only used as animal feed and fertilizer, while the rest is thrown away into the rivers or sea (Jelen, 2011).

A mozzarella producer in Bandung District, KPBS Pangalengan, was one of the producers that can’t turn whey into whey powder. One utilization of whey that has seen some popularity is the conversion into bioethanol and liquid organic fertilizer. This process can reduce the BOD of whey to 1,920 ppm, with emissions of CO2 to 13.65% (422,219.67 kg/CO2eq/year), and results in a positive community perception toward the environmental, social, and economic impacts (Utama et al., 2019). For further research, the nutritional component in whey has the potential to be utilized in more ways. That’s because, besides nutrition, whey also has some functional properties derived from amino acids and peptides.

Amino acids are protein monomers and peptides are components that contain two or more amino acids which are bound together by peptide bonds (Dullius et al., 2018). Essential amino acids in whey are higher than in eggs, casein, meat, and soybeans (Smithers, 2008). The functional properties of essential and nonessential amino acids are beneficial to human health, including the immune system, neurological system, anti-oxidative responses, protein synthesis, reproduction, and growth (Wu, 2010). Peptides in whey also have some functional properties such as antioxidant, antimicrobial, antihypertensive, anticancer, opioid, and immunomodulatory functions (Dullius et al., 2018). The presence of amino acids and peptides in whey can be enhanced by enzymatic reactions.

As an example of enzymatic reaction, fermentation needs several components such as a carbon source (Febrianti et al., 2017). Cheese whey has enough carbon with the presence of lactose, a component that is widely used in bioprocess media needing a lactic microorganism (Utama et al., 2017a). Fermentation by microorganisms has shown to be a cost-effective method and is widely used in the dairy industries (Daliri et al., 2017).

Fermentation of mozzarella whey can be accomplished with native yeast, because it lives naturally in mozzarella whey (Hossain et al., 2017). Six colonies were isolated from mozzarella cheese whey, three of them were identified as C. tropicalis and the rest were two isolates of Trichosporon beigelii, and one isolate of Blastoschizomyces capitatus (Balia et al., 2018). Therefore, C. tropicalis is the native yeast from mozzarella whey and has the potential to create the enzymatic reaction (Utba et al., 2018). C. tropicalis produces several proteinase and peptides can be produced by secretions of aspartyl proteinases families such as pepsin, cathepsin, and chymosin (Balia et al., 2018; Utba et al., 2018).

The fermentation time and the peptide-amino profile during fermentation need to be observed. The longer the fermentation, the more peptides will be converted into amino acids because of further enzymatic activity (Wang et al., 2017). The yeast released its proteolytic compounds into the protein material to discharge peptides and amino acids from the parent proteins (Daliri et al., 2017). The approximation of amino acids during protein hydrolysis didn’t allow for adequate evaluation of the concentration change to have the option to analyze hydrolysis and fermentation rates (Duong et al., 2019). Therefore, the research aimed to determine the optimal fermentation time in producing peptides and observed the amino acid profiles in native cheese whey before and after fermentation.


Conclusion

Natural cheese whey fermentation with C. tropicalis can produce maximum amounts of peptides in 24 hours. The amino acid profile after fermentation decreased all amino acids observed except isoleucine. Fermentation without the addition of C. tropicalis continued to increase and showed no peak in peptides until the 48-hour mark. The amino acid profile produced showed an increase in aspartate, glutamate, threonine, valine, isoleucine, and lysine and a decrease in serine, histidine, glycine, arginine, alanine, tyrosine, and methionine.

Acknowledgement

Authors would like to thank the Ministry of Research Technology and Higher Education that provided the research fund within the scheme of “Penelitian Tesis Magister” 2019 with the contract number of 1637/UN.6N/LT/2019, and The Rector of Universitas Padjadjaran for the Academic Leadership Grant. Authors would also to thank KPBS Pangalengan for kindly giving us permit for research, as well as Rudi Adi Saputra, Faysa Utba, Vivi Fadila Sari and Syarah Virgina for helping the authors in carrying out and obtaining research data.

References

Andrade, R.P., Melo, C.N., Genisheva, Z., Schwan, R.F., Duarte, W.F., 2017. Yeasts from Canastra Cheese Production Process: Isolation and Evaluation of Their Potential for Cheese Whey Fermentation. Food Research International, Volume 91, pp. 72–79

Balia, R.L., Kurnani, T.B.A., Utama, G.L., 2018. Selection of Mozzarella Cheese Whey Native Yeasts with Ethanol and Glucose Tolerance Ability. International Journal on Advanced Science, Engineering and Information Technology, Volume 8(4), pp. 1091–1097

Bartolomeo, M.P., Maisano, F., 2006. Validation of a Reversed-phase HPLC Method for Quantitative Amino Acid Analysis. Journal of Biomolecular Techniques, Volume 17(2), pp. 131–137

Belem, M.A.F., Gibbs, B.F., Lee, B.H., 1999. Proposing Sequences for Peptides Derived from Whey Fermentation with Potential Bioactive Sites. Journal of Dairy Science, Volume 82(3), pp. 486–493

Božani?, R., Baruk?i?, I., Lisak, K., Tratnik, L., 2014. Possibilities of Whey Utilisation. Austin Journal of Nutrition and Food Sciences, Volume 2(7), pp. 17

Chaves-López, C., Tofalo, R., Serio, A., Paparella, A., Sacchetti, G., Suzzi, G., 2012. Yeasts from Colombian Kumis as Source of Peptides with Angiotensin I Converting Enzyme (ACE) Inhibitory Activity in Milk. International Journal of Food Microbiology, Volume 159(1), pp. 39–46

Daliri, E.B.-M., Oh, D.H., Lee, B.H., 2017. Bioactive Peptides. Foods, Volume 6(5), pp. 127

Dullius, A., Goettert, M.I., de Souza, C.F.V., 2018. Whey Protein Hydrolysates as a Source of Bioactive Peptides for Functional Foods – Biotechnological Facilitation of Industrial Scale-up. Journal of Functional Foods, Volume 42, pp. 58–74

Duong, T.H., Grolle, K., Nga, T.T.V., Zeeman, G., Temmink, H., Eekert, M. van, 2019. Protein Hydrolysis and Fermentation Under Methanogenic and Acidifying Conditions. Biotechnol Biofuels, Volume 12, pp. 1–10

Febrianti, F., Syamsu, K., Rahayuningsih, M., 2017. Bioethanol Production from Tofu Waste by Simultaneous Saccharification and Fermentation (SSF) using Microbial Consortium. International Journal of Technology, Volume 8(5), p. 898–908

Han, B.-Z., Rombouts, F.M., Nout, M.J.R., 2004. Amino Acid Profiles of Sufu, A Chinese Fermented Soybean Food. Journal of Food Composition and Analysis, Volume 17 (6), pp. 689–698

Hawashi, M., Widjaja, T., Gunawan, S., 2019. Solid-State Fermentation of Cassava Products for Degradation of Anti-nutritional Value and Enrichment of Nutritional Value. In: New Advances on Fermentation Processes, Martínez-Espinosa, Rosa María (ed.)

Hossain, N., Haji Zaini, J., Mahlia, T.M.I., 2017. A Review of Bioethanol Production from Plant-based Waste Biomass by Yeast Fermentation. International Journal of Technology, Volume 8(1), pp. 518

Jelen, P., 2011. Whey Processing: Utilization and Products. In: Encyclopedia Dairy Science, John W. Fuquay (ed.), Academic Press, Mississippi, USA, pp. 2739–2745

KiBeom, L., Ho-Jin, K., Sang-Kyu, P., 2014. Amino Acids Analysis during Lactic Acid Fermentation by Single Strain Cultures of Lactobacilli and Mixed Culture Starter Made. African Journal Biotechnology, Volume 13(28), pp. 2867–2873

Korhonen, H., Pihlanto, A., 2003. Food-derived Bioactive Peptides - Opportunities for Designing Future Foods. Current Pharmaceutical Design, Volume 9(16), pp. 1297–1308

Liu, S.-Q., Holland, R., Crow, V.L., 2003. The Potential of Dairy Lactic Acid Bacteria to Metabolise Amino Acids Via Non-transaminating Reactions and Endogenous Transamination. International Journal of Food Microbiology, Volume 86(3), pp. 257–269

Panjaitan, F., Gomez, H., Chang, Y.-W., 2018. In Silico Analysis of Bioactive Peptides Released from Giant Grouper (Epinephelus lanceolatus) Roe Proteins Identified by Proteomics Approach. Molecules, Volume 23(11), pp. 115

Riceto, É.B. de M., Menezes, R. de P., Penatti, M.P.A., Pedroso, R. dos S., 2015. Enzymatic and Hemolytic Activity in Different Candida Species. Revista Iberoamericana de Micología, Volume 32(2), pp. 79–82

Rochín-Medina, J.J., Ramírez-Medina, H.K., Rangel-Peraza, J.G., Pineda-Hidalgo, K.V., Iribe-Arellano, P., 2018. Use of Whey as a Culture Medium for Bacillus clausii for the Production of Protein Hydrolysates with Antimicrobial and Antioxidant Activity. Food Science and Technology International, Volume 24(1), pp. 35–42

Shah, A.H., Hameed, A., Khan, G.M., 2002. Fermentative Production of L-Lysine: Bacterial Fermentation-I. Journal of Medical Sciences, Volume 2(3), pp. 152–157

Smithers, G.W., 2008. Whey and Whey Proteins—from ‘gutter-to-gold.’ International Dairy Journal, Volume 18(7), pp. 695–704

Suryaningsih, V., Ferniah, R.S., Kusdiyantini, E., 2018. Karakteristik Morfologi, Biokimia, dan Molekuler Isolat Khamir Ik-2 Hasil Isolasi dari Jus Buah Sirsak, Annona muricata L. (Morphological, Biochemical, and Molecular Characteristics of Yeast Ik-2 Isolated from Soursop Fruit Juice). Jurnal Biologi, Volume 7(1), pp. 18–25

Utama, G.L., Januaramadhan, I., Dinika, I., Balia, R.L., 2019. Bioconversions of Cheese-Making Wastes to Bioethanol and Their Link to Sustainability. In: IOP Conferences Series: Earth Environmental Science, Volume 306, pp. 18

Utama, G.L., Kurnani, T.B.A., Sunardi, Balia, R.L., 2017a. Reducing Cheese-making By-product Disposal through Ethanol Fermentation and the Utilization of Distillery Waste for Fertilizer. International Journal of GEOMATE, Volume 13(37), pp. 103–107

Utama, G.L., Kurnani, T.B.A., Sunardi, S., Cahyandito, F., Balia, R.L., 2017b. Joint Cost Allocation of Cheese-making Wastes Bioconversions into Ethanol and Organic Liquid Fertilizer. Bulgarian Journal of Agricultural Science, Volume 23, pp. 1016–1020

Utba, F., Balia, R.L., Utama, G.L., 2018. The Presence of Indigenous Yeasts with Proteolytic Activity Isolated from Homemade-Mozzarella Whey. Scientific Papers Series Management, Economic Engineering in Agriculture and Rural Development, Volume 18(1), pp. 507–514

Wang, W., Xia, W., Gao, P., Xu, Y., Jiang, Q., 2017. Proteolysis during fermentation of Suanyu as a Traditional Fermented Fish Product of China. International Journal of Food Properties, Volume 20(sup1), pp. S166–S176

Wu, G., 2010. Functional Amino Acids in Growth, Reproduction, and Health. Advances in Nutrition, Volume 1(1), pp. 31–37