Biopulping Of Bagasse Using Different Types Of White Rot Fungi and Different Incubation Times

Rekam Jejak Artikel: Diterima : 30/08/2019 Disetujui : 19/11/2020 Abstract Bagasse is fibrous residue that remains after the sugarcane is crushed to extract it’s juice. It mostly consists of lignocellulosic materials that may provide material for paper production through biopulping process. White rot fungi (WRF) is producers of extracellular ligninolytic enzymes that has the capability to mineralize lignin compounds. Three types of white rot fungi that were used in this study are Phanerochaete chrysosporium, Pleurotus ostreatus and Schizophyllum commune. Each of it was inoculated on bagasse substrate within 0, 15, and 30 days of incubation. This study was aimed to know the effect of interaction between white rot fungi and incubation time in the biopulping process and to investigate the most appropriate fungus and incubation time to produce good material for paper making obtained from sugarcane bagasse. The experimental design was done by using Completely Randomized Design (CRD) with a factorial pattern in two factors and analyzed by using Analysis of Variance (ANNOVA) then followed by Duncan’s Multiple Range Test (DMRT) on highly different significance effect of the treatment. The result of this study showed that S. commune is the most effective fungi to degrade highly lignin content (17.38% to 8.88%) at 30 days of incubation, while P. chrysosporium is the most effective fungi to lowering cellulose content in small amount (23.64% to 19.38%) during 30 days of incubation.


INTRODUCTION
Central and East Java are centers of cane sugar industry in Indonesia because the area of sugarcane plantation in this area is wide. According to Badan Pusat Statistik (2015), the largest sugarcane production derived from East Java Province which is amounted to 1.24 million tons, whereas the sugarcane production that is located in Central Java is amounted to 206.25 thousand tons.
The production of sugar cane plantation always remains sugarcane leaves and bagasse in large amount (Khuluq, 2012).
Bagasse is the fibrous residue that remains after the sugarcane is extracted into it's juice and mostly contains of lignocellulose compound such as lignin (13-22%), cellulose (26-43%), hemicellulose (17-23%) and pentosan (20-33%) (Johnson, & Vikram, 2012). The utilization of bagasse will provide more benefits in reducing deforestation rather and can be used as a raw materials for pulp and paper making (Al-Sulaimani & Dwivedi, 2017). Bagasse is derived from the sugar cane plant (Saccharum officinarum) which grows in tropical and sub tropical areas. It is a member of family Gramineae which has morphology of stem height approximately 3-5 m, does not have branches and grows upright. The fiber of bagasse is insoluble in water and consists mainly of cellulose, pentosan and lignin (Rulianah et al., 2017). The process of degrading lignin is necessary to do by lignin-degrading fungi before making pulp through biopulping process.
Biopulping is the treatment of lignocellulose degradation with a fungus which have ligninolytic enzyme and the subsequent processing of material can be done through mechanical or chemical pulping (Saad et al., 2008). The best treatment of the biopulping process is treatment which results in a high decrease of lignin content but minimal loss of cellulose content. The concept of biopulping is based on the ability of fungi to colonize and degrade lignin selectively in wood thereby remains cellulose relatively intact (Singhal et al., 2015).
One part of biopulping is biodelignification.
Biodelignification is a process which use biological agents or organisms that are capable of releasing cellulosic fibers from a lignin bonds by its natural ability (Chang & Miles, 1989).
White rot fungi (WRF) which is classified in the Basidiomycota group is the most effective organisms that able to degrade the whole wood components due to the secretion of extracellular ligninolytic enzymes such as lignin peroxidase (LiP), manganese dependent peroxidase triplicate (MnP), and laccase that has the capability to mineralize lignin and phenolic organic substrates.
According to Musa et al. (2012), White rot fungi will break down lignin completely into water (H 2 O) and carbon dioxide (CO 2 ) At the end process of lignin degrading, they will leave decayed wood by whitish in color and fibrous in texture. They are preferentially attacking lignin more readily and This situation can occurs because the incubation time will give the opportunity to fungi to use organic materials as an energy source and carbon source to form new cells and produce ligninbreaking enzymes (Chang & Miles, 1989).
Biopulping process requires longer incubation time for growth of fungal biomass and delignification. However, the longer the incubation time the higher losses of bagasse chemical components such as extractive substances, lignin, cellulose and hemicellulose can be occured. It may be caused by differences capability of each fungi in the production of enzyme to degrade the substrate.
According to Musa et al. (2012), P. (2) and to investigate the most appropriate fungus and incubation time to produce good material for paper making from sugarcane bagasse.

Materials
The materials were used are isolate of

Research Design
The experiment was conducted using

Sample Preparation
Bagasse sample was strained through a muslin cloth to remove from dirt and then dried under sunlight for approximately 5 hours in a day until dry. Further, the bagasse was collected into a cloth carry bag.

Fungal Spawn Inoculum Preparation (Kusnadi & Yayan, 2003, modified)
Sorghum as much as 5 kg was prepared. It was soaked in water for approximately 2 hours.
Floating dirt and seeds were removed. The sorghum was drained and then steamed with water until the seeds burst at one point.

Preparation of Bagasse Substrate (Sidana & Umar, 2014, modified)
Bagasse which has been processed was take out from cloth carry bag, drained and then put it into a container. The distilled water as much as 1L and bagasse as much as 1 kg were prepared. The

Analysis of Lignin and Cellulose content (Chesson, 1981)
Analysis of lignin and cellulose before and after incubation time (0, 15, and 30 days) was done by following the Chesson method

Analysis Data (Weber & Skillings, 2000, modified)
The quantitative data of the lignin and cellulose contents at the beginning and the end of treatment were analyzed by using analysis of variance (ANNOVA) with 5% significant. Then it was followed by Duncan's Multiple Range Test (DMRT) to determine the difference between each treatment.

Lignin Contents
The content of lignin on bagasse substrate can be degraded by using the treatment of    The least average reduction of lignin content can be showed by the treatment of P.
chyrsosporium fungi. In this study, the growth of P.
chrysosporium fungi was slower compared to other fungi, S. commune and P. ostreatus fungi.
According to Herliyana (1997), this was thought due to several factors, such as the origin of isolates and the media growth that were used.

Cellulose Content
The content of cellulose on bagasse substrate also can be degraded by using the treatment of P. chrysosporium, P. ostreatus, and S.  Therefore, this cellulose content can be used as the initial cellulose content. fungi is one type of non-selective fungi which had fast growth and can causes a weathering and wood discoloration. This fungus also had the enzyme of selobiohydrolase and endo1.4 β-glucanase for cellulose degradation.
According to Rodríguez-Couto (2017), during pulp and paper production, it is necessary to separate the cellulose fibers from lignin.
Biopulping is a good treatment for wood chips processing with ligninolytic fungi prior to conventional pulping methods either mechanical or chemical pulping methods. The benefits of biopulping in the production of paper were environmentally-friendly, cheap cost and can reduce not only energy consumption but also the utilization of chemicals.