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Identification of Bacterial Isolates from Mozzarella Cheese Whey Using 16S rDNA and Assessment of Their Consortium Proteolytic Activity: Identification and Protease Activity of Cheese Whey Bacteria

Journal of Tropical Life Science : International Journal of Theoretical, Experimental, and Applied Life Sciences · Vol. 15 No. 1 (2025) · hlm. 153–160

DOI
10.11594/jtls.15.01.15
ISSN
2527-4376, 2087-5517
Terbit
2025-06-26
Penerbit
Journal of Tropical Life Science
Bahasa
eng
Lisensi
creativecommons.org/licenses/by-nc/4.0

Abstrak

Cheese whey is the main waste from cheese production, which contains high levels of protein, lactose, and minerals. However, if not properly managed, its acidic pH and high organic load can negatively impact the environment when discharged directly. Its protein content makes it a habitat for proteolytic bacteria that are capable of producing protease enzymes to hydrolyze proteins into peptides and amino acids. Moreover, the utilization of proteolytic bacteria from cheese whey offers a biotechnological solution for waste management while opening up opportunities for industrial applications, such as fermentation and the production of functional enzymes. The aim of this study is to analyze the protease activity of single bacterial isolates and bacterial consortia isolated from cheese whey and identify isolates based on 16S rDNA. The first stage of the research was screening pathogenicity based on hemolysis analysis using a blood agar medium. The second stage tested for synergism among isolates. The ability of isolates to produce proteolytic enzymes qualitatively and quantitatively, and then the identification of proteolytic bacterial species based on 16S rDNA. Hemolysis assay of eight isolates resulted in three isolates (A8, C7, and C8) showing gamma hemolysis. The protease activity assay of three isolates was measured at an incubation period of 0-72 hours; the consortium (cn) isolate exhibited the highest activity of 0.47 U/mL U/mL. Based on the 16S rDNA sequence, isolate A8 was identified as Staphylococcus saprophyticus 36QC2CO with a similarity of 98.04%, isolate C7 was identified as S. saprophyticus WWi54 with a similarity of 97.81%, and isolate C8 was identified as Staphylococcus epidermidis 2322 with a similarity of 97.6%.

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  • University of Brawijaya
  • Universitas Maarif Hasyim Latif
  • North Carolina Agricultural and Technical State University
  • Universitas Cokroaminoto Palopo
  • Universitas Katolik Widya Mandira
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Rekan penulisOrganisasiArtikel bersama
Muhaimin Rifa’iUniversity of Brawijaya3
Irfan MustafaUniversity of Brawijaya2
Nashi WidodoUniversity of Brawijaya2
Dwi Purno WidekdoUniversity of Brawijaya2
Agung Putra PurwantoUniversity of Brawijaya2
Alifia Issabella MulyawatiUniversity of Brawijaya2
Elsa Mega SuryaniUniversitas Maarif Hasyim Latif1
Hikmah MaulidiyahUniversity of Brawijaya1
Salam Adnan IbrahimNorth Carolina Agricultural and Technical State University1
Suharjono SuharjonoUniversity of Brawijaya1
Elok ZubaidahUniversity of Brawijaya1
Eka Pratiwi TenriawaruUniversitas Cokroaminoto Palopo1
Endang ArisoesilaningsihUniversity of Brawijaya1
Eufrasia R. A. LengurUniversitas Katolik Widya Mandira1
Eko WidodoUniversity of Brawijaya1
Farida RachmawatiUniversity of Brawijaya1
Sri WidyartiUniversity of Brawijaya1
Aris SoewondoUniversity of Brawijaya1
Sri RahayuUniversity of Brawijaya1
Fikriya Novita SariUniversity of Brawijaya1
Sefihara Paramitha SaktiUniversity of Brawijaya1
Osfar SjofjanUniversity of Brawijaya1
Sulistya Ika RamadhaniAfiliasi tidak diketahui1
Aleyda WahyudiAfiliasi tidak diketahui1
Giral Baines MoraUniversity of Brawijaya1
Dewi PermatasariUniversity of Brawijaya1
Maharani Pertiwi KoentjoroUniversity of Brawijaya1
Syarafin MahiraUniversity of Brawijaya1
asep awaludin prihantoAfiliasi tidak diketahui1
Yoga Dwi JatmikoAfiliasi tidak diketahui1
Rika Gian MeliantiAfiliasi tidak diketahui1
Asep Awaludin PrihantoUniversity of Brawijaya1
Rika Gian MeliantiUniversity of Brawijaya1

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Sumber metadata: OAI-PMH (metadata jurnal) http://jtrolis.ub.ac.id/index.php/jtrolis/oai, pertama di-harvest 9 Nov 2026, terakhir terlihat 9 Nov 2026.
Metadata asli (oai_dc) 
<oai_dc:dc xmlns="http://www.openarchives.org/OAI/2.0/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:oai_dc="http://www.openarchives.org/OAI/2.0/oai_dc/" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xsi:schemaLocation="http://www.openarchives.org/OAI/2.0/oai_dc/  http://www.openarchives.org/OAI/2.0/oai_dc.xsd">
	<dc:title xml:lang="en">Identification of Bacterial Isolates from Mozzarella Cheese Whey Using 16S rDNA and Assessment of Their Consortium Proteolytic Activity: Identification and Protease Activity of Cheese Whey Bacteria</dc:title>
	<dc:creator xml:lang="en">Maulidiyah, Nuris</dc:creator>
	<dc:creator xml:lang="en">Ardyati, Tri </dc:creator>
	<dc:creator xml:lang="en">Dwi Jatmiko, Yoga</dc:creator>
	<dc:subject xml:lang="en">Cheese whey</dc:subject>
	<dc:subject xml:lang="en">Identification of 16S rDNA</dc:subject>
	<dc:subject xml:lang="en">Proteolytic bacteria</dc:subject>
	<dc:description xml:lang="en">Cheese whey is the main waste from cheese production, which contains high levels of protein, lactose, and minerals. However, if not properly managed, its acidic pH and high organic load can negatively impact the environment when discharged directly. Its protein content makes it a habitat for proteolytic bacteria that are capable of producing protease enzymes to hydrolyze proteins into peptides and amino acids. Moreover, the utilization of proteolytic bacteria from cheese whey offers a biotechnological solution for waste management while opening up opportunities for industrial applications, such as fermentation and the production of functional enzymes. The aim of this study is to analyze the protease activity of single bacterial isolates and bacterial consortia isolated from cheese whey and identify isolates based on 16S rDNA. The first stage of the research was screening pathogenicity based on hemolysis analysis using a blood agar medium. The second stage tested for synergism among isolates. The ability of isolates to produce proteolytic enzymes qualitatively and quantitatively, and then the identification of proteolytic bacterial species based on 16S rDNA. Hemolysis assay of eight isolates resulted in three isolates (A8, C7, and C8) showing gamma hemolysis. The protease activity assay of three isolates was measured at an incubation period of 0-72 hours; the consortium (cn) isolate exhibited the highest activity of 0.47 U/mL U/mL. Based on the 16S rDNA sequence, isolate A8 was identified as Staphylococcus saprophyticus 36QC2CO with a similarity of 98.04%, isolate C7 was identified as S. saprophyticus WWi54 with a similarity of 97.81%, and isolate C8 was identified as Staphylococcus epidermidis 2322 with a similarity of 97.6%.</dc:description>
	<dc:publisher xml:lang="en">Journal of Tropical Life Science</dc:publisher>
	<dc:date>2025-06-26</dc:date>
	<dc:type>info:eu-repo/semantics/article</dc:type>
	<dc:type>info:eu-repo/semantics/publishedVersion</dc:type>
	<dc:format>application/pdf</dc:format>
	<dc:identifier>https://jtrolis.ub.ac.id/index.php/jtrolis/article/view/3629</dc:identifier>
	<dc:identifier>10.11594/jtls.15.01.15</dc:identifier>
	<dc:source xml:lang="en">Journal of Tropical Life Science; Vol. 15 No. 1 (2025); 153 – 160</dc:source>
	<dc:source>2527-4376</dc:source>
	<dc:source>2087-5517</dc:source>
	<dc:language>eng</dc:language>
	<dc:relation>https://jtrolis.ub.ac.id/index.php/jtrolis/article/view/3629/788</dc:relation>
	<dc:rights xml:lang="en">Copyright (c) 2025 Journal of Tropical Life Science</dc:rights>
	<dc:rights xml:lang="en">https://creativecommons.org/licenses/by-nc/4.0</dc:rights>
</oai_dc:dc>