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Screening of Cellulolytic Bacteria from Sugarcane Waste (Bagasse) and Optimization of Cellulase Activity as Animal Feed: Screening of Cellulolytic Bacteria from Sugarcane Waste (Bagasse) Ramadhani, Sulistya Ika; Ardyati, Tri; Sjofjan, Osfar
Journal of Tropical Life Science Vol. 13 No. 3 (2023)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.13.03.19

Abstract

The Sugar Factory of Tjoekir Jombang is one of the sugar factories located in Jombang, East Java under PT Perkebunan Nusantara X. Sugarcane waste, also known as bagasse causes pollution of the environment. Some microorganisms are able to degrade cellulose-containing bagasse, because of cellulolytic enzymes produced by cellulolytic bacteria. This study aims to obtain cellulolytic bacteria isolates, screen the highest cellulolytic activity, identify the highest cellulolytic activity isolates, and optimize conditions (pH and temperature) for the highest cellulase activity. Cellulolytic bacteria from bagasse were grown on a medium containing 1% CMC. Several additional minerals were other than those in the CMC medium. The cellulase activity was assayed semi-quantitatively with the addition of 0.1% Congo red and quantitatively using the 3,5-Dinitro Salicylic Acid (DNS) method. Bacterial isolates with high cellulolytic activity were identified based on the 16S rDNA sequence. This research obtained 20 bacterial isolates, where isolate A1T4 had the highest cellulolytic index of 1.18 mm. Measurement of cellulase activity using the DNS method showed that isolate A2T2 had the highest cellulase activity of 2.19 U/mL. Hemolysis assay showed that from 12 isolates, only two isolates have γ-hemolysis activity (isolates A1T6 and A3T3). Those isolates were optimized in a CMC broth medium with temperatures of 30°C and 37°C and pH 5 and pH 6 to produce the highest cellulase activity. Isolate A1T6 and A3T3 were grown optimally at 30oC and pH 6. Isolate A1T6 was identified as Citrobacter amalonaticus with a similarity of 99.80%, and isolate A3T3 was identified as Pseudomonas mendocina with a similarity of 98.83%.
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 Maulidiyah, Nuris; Ardyati, Tri; Dwi Jatmiko, Yoga
Journal of Tropical Life Science Vol. 15 No. 1 (2025)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.15.01.15

Abstract

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%.
Screening of Cellulolytic Bacteria from Sugarcane Waste (Bagasse) and Optimization of Cellulase Activity as Animal Feed: Screening of Cellulolytic Bacteria from Sugarcane Waste (Bagasse) Ramadhani, Sulistya Ika; Ardyati, Tri; Sjofjan, Osfar
Journal of Tropical Life Science Vol. 13 No. 3 (2023)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.13.03.19

Abstract

The Sugar Factory of Tjoekir Jombang is one of the sugar factories located in Jombang, East Java under PT Perkebunan Nusantara X. Sugarcane waste, also known as bagasse causes pollution of the environment. Some microorganisms are able to degrade cellulose-containing bagasse, because of cellulolytic enzymes produced by cellulolytic bacteria. This study aims to obtain cellulolytic bacteria isolates, screen the highest cellulolytic activity, identify the highest cellulolytic activity isolates, and optimize conditions (pH and temperature) for the highest cellulase activity. Cellulolytic bacteria from bagasse were grown on a medium containing 1% CMC. Several additional minerals were other than those in the CMC medium. The cellulase activity was assayed semi-quantitatively with the addition of 0.1% Congo red and quantitatively using the 3,5-Dinitro Salicylic Acid (DNS) method. Bacterial isolates with high cellulolytic activity were identified based on the 16S rDNA sequence. This research obtained 20 bacterial isolates, where isolate A1T4 had the highest cellulolytic index of 1.18 mm. Measurement of cellulase activity using the DNS method showed that isolate A2T2 had the highest cellulase activity of 2.19 U/mL. Hemolysis assay showed that from 12 isolates, only two isolates have γ-hemolysis activity (isolates A1T6 and A3T3). Those isolates were optimized in a CMC broth medium with temperatures of 30°C and 37°C and pH 5 and pH 6 to produce the highest cellulase activity. Isolate A1T6 and A3T3 were grown optimally at 30oC and pH 6. Isolate A1T6 was identified as Citrobacter amalonaticus with a similarity of 99.80%, and isolate A3T3 was identified as Pseudomonas mendocina with a similarity of 98.83%.