Bio Pulping Of Bagasse As The Material For Paper Making Using Different Species Of White Rot Fungi And Incubation Time

Rekam Jejak Artikel: Diterima : 28/08/2019 Disetujui : 28/11/2020 Abstract The Biopulping is defined as the biological process of lignin degradation by utilizing microorganisms that produce some enzymes. A microorganism which widely known in the degradation of lignin and cellulose is a group of white-rot fungi. The aims for this research to know the most effective white rot fungi species of G.lucidum, P.tuber-regium, and T.versicolor in the degradation of lignin and cellulose with different incubation time on bagasse substrate. The effectivity of biopulping indicated by the highest degradation of lignin concentration and the lowest degradation of cellulose concentration. This study used an experimental design with Completely Randomized design with a two factorial pattern. The independent variable of this study is white rot fungi species and incubation time while the dependent variable is the concentration of lignin and cellulose. The main parameter was the concentration of lignin and cellulose, supporting parameters were pH, weight loss of substrate and mycelial growth. The result showed the degradation of lignin and cellulose in all treatment. The best degradation of lignin and cellulose showed by species T.versicolor and P.tuber-regium within 30 days of incubation. Keyword: Bagasse; Biopulping; White Rot Fung

The waste of sugarcane mostly used as bioethanol.
Bagasse and straw are mostly used for generating heat and electricity, and recently bagasse is used as paper material. The process of papermaking from bagasse can be done through biopulping (Del Rio et al., 2015).
Bio-pulping is an environmentally friendly technology which currently being developed in the pulp and paper industry. The application of biopulping in the pulp and paper industry is before the cooking process using chemicals. The pulping process pretreated with White Rot Fungi (WRF) has proven as a technology that is efficient and economically feasible, energy savings, reducing chlorine consumption and reducing pollutants into the environment and produce stronger pulp.

Materials
The materials used were G. lucidum, P.

Design of Research
The experimental design used in this research was Completely Randomized Design (CRD) with a Factorial pattern with 3 replication.
The independent variables were three species of white rot fungi and incubation time, while the dependent variable was the degradation of lignin and cellulose. The main parameters were concentration of lignin and cellulose before and after treatment. The supporting parameters were pH of substrate and weight loss of bagasse before and after treatment, and mycelial growth.

The procedure of Research
The laboratory equipment was used in this research such as petri dish, reaction tubes, beaker glass, measuring glass, and other tools were sterilized using autoclave with temperature 121ᵒC, 2 atm, for 15 minutes (Hadioetomo, 1994)  The isolates of G. lucidum, P. tuber-regium, and T.versicolor in PDA medium prepared.
Inoculation done by forming a plug using cork borer, and take as much as five plugs of each fungal and take it into a bottle medium and wrapped the medium aseptically in LAF then incubated it for one month.

Carolina, 2017, modified).
Bagasse substrate was taken 1 kg and put into a container, then 1 L distilled water was measured and put it container and homogenized.
The pH of the bagasse also measured using the soil tester. The bagasse substrate filled as much 200 g into 25x20x0.008 cm polypropylene plastic bags.
The baglogs were carried out with the process of compaction. The paralon ring was given on the top of the baglog followed by putting the cotton and plastic. The bagasse substrate medium was sterilized in the autoclave for 2 hours at 121 o C. The media that has been sterilized was cooled for 12 hours before inoculation.

Fungal Inoculation on Bagasse (Suryani & Carolina, 2017, modified).
The fungi isolated from sorghum medium was taken 10% from the weight of the bagasse substrate medium. Inoculation was done by making a deep hole approximately 2 cm in the middle of the baglog media using the sterilized wooden stick.
Then fungal spawn was inserted into the polybag by using spatula aseptically. It's incubated in room temperature with an incubation time of 0, 15, and 30 days.

Measurement pH in Bagasse Substrate
The measurement of pH in the bagasse substrate was done with soil tester tools. The soil tester was dipped into bagasse medium and the pH measured. The measurement was done before the treatment and after treatment.

Measurement of Weight Loss Media
The weight loss measurement of bagasse substrate has been done by measuring the bagasse substrate on day 0, 15 days and 30 days before and after treatment using an analytical scale.

Lignin and Cellulose Analysis (Chesson, 1981)
Determine lignin and cellulose concentration with different incubation time (0, 15, and 30 days) was done by Chesson method.

Data Analysis
The quantitative data of the lignin and cellulose contents at the beginning and the end of treatments were analyzed using analysis of variance (F test) with the error contents is 5% and 1% followed by Duncan's Multiple Range Test (DMRT).

Lignin Concentration
The degradation of lignin concentration in bagasse using three species of white-rot fungi were occurred in all treatment (

Cellulose Concentration
The degradation of cellulose concentration in bagasse using three species of WRF and different incubation time also showed the degradation in cellulose concentration. The initial cellulose concentration of bagasse was 23.64%.    The initial pH was 6.8. After 15 days of incubation the average pH of G. lucidum, P.tuberregium and T.versicolor ranged between 6.5 -6.6 while the treatment of 30 days incubation showed that the average pH was 5.43 -5.7 (Table 4). These pH values are suitable for the growth of the fungi.
Generally, the value of pH for fungal growth ranges between 5-6. According to Away et al. (2000), to utilize hemicellulose, cellulose and  Table 4