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Background: The record and research on macrofungi from Taman Eden 100, Toba-Samosir Regency, North Sumatra, Indonesia, was conducted to provide some information about their current status and potential use in the future. Methods: This research was divided into three steps: exploration, identification, and literature studies of the potential use from the identified macrofungi. Results: The research had successfully identified 14 macrofungi that were classified into 4 orders and 9 families. All the macrofungi found were Basidiomycota, namely Gymnopilus sp., Marasmiellus sp.1, Marasmiellus sp.2, Marasmius sp., Favolaschia sp., Coprinellus sp., Coprinopsis sp., Auricularia sp.1, Auricularia sp.2, Auricularia sp.3, Auricularia sp.4, Tylopilus sp., Suillus sp., and Russula sp. The identified macrofungi are potentially used as food supplements, medicine, bio-fertilisers, bioherbicides, and bioremediation agents based on the literature. Conclusions: Taman Eden 100 has a unique fungal diversity of macrofungi and has never been published in a scientific journal. Further investigations are needed to determine the fungal diversity and potential use of macrofungi in other places in Taman Eden 100.

Water pollution by dyes represents from dyestuff industry becomes an environmental concern. Finding new isolates capable of decolorizing these dyes is important. The study aimed to assess the new isolates of white-rot fungi (WRF) as decolorizing agent of anthraquinone and azo dyes. Decolorization assay were conducted in Agar plates and liquid medium. During the decolorization, laccase activities produced by the fungal strains were analyzed. Identification of the fungal strains were investigated using molecular DNA analysis. The results showed that isolates M3, H18, and GP1 were able to decolorize anthraquinone and azo dyes in Agar and liquid medium. Based on DNA analysis, isolates M3, H18, and GP1 have the similarity to Trametes sanguinea, Trametes polyzona, and Neofomitella guangxiensis, respectively. Among the fungi, T. polyzona H18 exhibited high decolorization ability (70–90%) to the dyes (100 mg/L) after 96-hours incubation. Laccase activity was fluctuated during the reactions with tendency to increase at the beginning until its peak, then decreased at the end of incubation. This study demonstrated the potential of the new isolates from Indonesia to decolorize anthraquinone and azo dyes. The results of the study can provide an alteranative for bioremediation agents of contaminated water by synthetic dyes.

Oil palm empty fruit bunch (OPEFB) has high potency to be converted into second generation bioethanol production. OPEFB pretreated liquid hot water (LHW) assisted maleic acid (MA) pretreatment was used as substrate in simultaneous saccharification fermentation (SSF). This pretreatment was conducted at 180 °C for 70 min by using 1.5% (w/v) MA acid concentration with material and liquid ratio of 1:14. Subsequently, the pretreated samples were fermented by addition of surfactant, polyethylene glycol (PEG) 4000 and amphiphilic lignin derivatives (A-LD) using Kluyveromyces marxianus InaCC Y119 yeast at temperature of 38 °C for 89 h. Before SSF, pretreated OPEFB was pre hydrolyzed by cellulase enzyme with enzyme loading 30 FPU/g at 50 °C for 3 h. Fermentation filtrate was taken at 0, 24, 48, 72, dan 89 h as much as 1.5 mL for reducing sugar and ethanol content determination. The surfactant addition can enhance reducing sugar and also ethanol content compared to control. The highest ethanol content of 1.251 g/L was found at fermentation of OPEFB pretreated MA with PEG 4000 surfactant at 89 h.

Recently, the interest in active packaging utilization has increased with population growth, food demand and new consumer trend like food delivery services. This new system, however, requires the use of additives to extend the food product quality and safety as well as in maintaining the shelf-life. This study was to prepare the antimicrobial paper from I. cylindrica coated anionic nanocellulose crosslinked cationic to create a system with the ability to actively control microbe growth in the packaging materials. The process involved pulping of I. cylindrica using semi-chemical and soda chemical method. The antimicrobial paper was prepared by printing the pulp suspension in 60 g/m2 grammage in mold followed by the spray of anionic nanocellulose and subsequent soaking of the paper in cationic solution. The results showed the I. cylindrica paper coated anionic nanocellulose crosslinked with H+ and Al3+ cations were successfully produced. The paper produced was also observed to have antimicrobial activity against Gram-negative of E. coli and S. typhi as well as Gram-positive of S. aureus and B. subtilis bacteria. Furthermore, the best coating method was found on antimicrobial paper coated anionic nanocellulose crosslinked Al3+ as evidenced by smoother and compact surface structure.

Upscaling of biomass pretreatment from laboratory scale to a bench-scale reactor is one of the important steps in the application of the pretreatment for pilot or commercial scale. This study reports the optimization of pretreatment conditions, namely reaction temperature and time, by one factor at a time (OFAT) method for the enhancement of enzymatic saccharification of oil palm empty fruit bunch (OPEFB). OPEFB was pretreated using high temperature-pressure steam reactor with different reaction temperatures (160, 170, 180, 190, 200 °C) and times (10, 20, 30, 40, 50 min). The effectiveness of the pretreatment was determined based on chemical compositions of raw OPEFB and OPEFB pulp and sugar production from enzymatic saccharification of the OPEFB pulp . Solubilized components from OPEFB, such as glucose, xylose, formic acid, acetic acid, 5-hydroxymethyl furfural (HMF), and furfural in the hydrolysate that generated during steam pretreatment were also determined. Pretreatment at 180 °C for 20 min provide s the highest sugar yields (97.30% of glucose yield per initial cellulose and 88.86% of xylose yield per initial hemicellulose). At the optimum condition, 34.9% of lignin and 30.75% of hemicellulose are successfully removed from the OPEFB and resulted in 3.43 delignification selectivity. The relationship between severity factor and by-products generated and the sugars obtained after enzymatic saccharification are discussed. The pulp of OPEFB at the optimum condition was also characterized for its morphological characteristic by scanning electron microscopy (SEM) and crystallinity by X-ray diffractometry (XRD) . These pulp characteristics are then compared with those of the raw OPEFB . The steam pretreatment cause s some fiber disruptions with more defined and opened structures and increase s the crystallinity index (CrI) by 2.9% compared to the raw OPEFB.

Hydrolysis of biomass for the production of fermentable sugar can be improved by the addition of surfactants. In pulp and paper mills, lignin, which is a by-product of the pulping process, can be utilized as a fine chemical. In the hydrolysis process, lignin is one of the major inhibitors of the enzymatic breakdown cellulose into sugar monomer. Therefore, the conversion of lignin into a biosurfactant offers the opportunity to solve the waste problem and improve hydrolysis efficiency. In this study, lignin derivatives, a biosurfactant, was applied to enzymatic hydrolysis of various lignocellulosic biomass. This Biosurfactant can be prepared by reacting lignin with a hydrophilic polymer such as polyethylene glycol diglycidylethers (PEDGE). In this study, the effect of biosurfactants on the enzymatic hydrolysis of pretreated sweet sorghum bagasse (SSB), oil palm empty fruit bunch, and sugarcane trash with different lignin contents was investigated. The results show that lignin derivatives improve the enzymatic hydrolysis of the pretreated biomass with low lignin content, however, it has less influence on the enzymatic hydrolysis of other pretreated biomass with lignin content higher than 10% (w/w). The use of biosurfactant on SSB kraft pulp can increase the sugar yield from 45.57% to 81.49%.

Nurhayat OD, Ardiati FC, Ramadhan KP, Anita SH, Okano H, Watanabe T, Yanto DHY. 2021. Bioprospecting three newly isolated white-rot fungi from Berbak-Sembilang National Park, Indonesia for biodecolorization of anthraquinone and azo dyes. Biodiversitas 22: 613-623. Extensive use of textile dyes without proper wastewater treatment may jeopardize the water environment. In this study, bioprospecting newly isolated white-rot fungi from Berbak-Sembilang National Park for decolorizing four synthetic dyes was investigated. A total of 108 wood-decaying fungi were screened by using selective media, resulting in three isolates as the most promising fungal strains (BRB 11, BRB 73, BRB 81). BRB 81 had the best ability to decolorize 91.4% of AB129 and 77.8% of RB5 within 96 hours while the highest removal of RBBR and AO7 was performed by BRB11 around 60% and 37.6%, respectively. The enzymatic degradation was assumed to involve the decolorization process as laccase activities were observed with the highest around 116 UL-1. Based on molecular identification, these three fungal isolates were identified as Phellinus noxius BRB 11, Ceriporia lacerata BRB 81, and Leiotrametes menziesii BRB 73, respectively. In conclusion, P. noxius BRB 11, L. menziesii BRB 73, and C. lacerata BRB 81 could be used as biological agents in textile wastewater treatment and thus, it is important to conserve them as a part of the biodiversity within the local biosphere reserve.

Crude laccase from Trametes hirsuta EDN 082 was used in this study for the decolorization of anthraquinone and azo dyes. Characterization of degradation products from one selected dye, Acid Blue 129 (AB129), was investigated using liquid chromatography high-resolution mass spectrometry (LC-HRMS). The results showed that laccase from T. hirsuta EDN 082 (0.25 U/mL dyes) was able to decolourize AB 129, AB 113, RBBR, AB 25, RB4, and RB5 dyes with a decolorization efficiency of 88%, 73%, 71%, 72%, 42%, and 6%, respectively for 7 h. During the degradation of AB129 dye, nine metabolites were identified and characterized by electrospray ionization–liquid chromatography–mass spectrometry Quadrupole-Time of flight (ESI-LCMS-Q-TOF) analysis. Evaluation of peak area showed decreasing of the two metabolites, 1-amino-4-hydroxy-9,10-dioxo-9,10-dihydroanthracene-2-sulfonic acid (D2) and 2,4,6-trimethylaniline (D4) during 7 h reaction indicated these two compounds become the centre of the degradation after cleaving the AB129 dye, and from these results, metabolic pathways were proposed. This study offers the potency of laccase from T. hirsuta EDN 082 for bioremediation of synthetic dyes in the fields.

Pretreatment is an important step in the conversion of biomass to bioethanol. In this study microwave-assisted oxalic acid (MOA) pretreatment was chosen to pretreat oil palm empty fruit bunch (OPEFB) to enhance enzymatic saccharification of the biomass. The objective of this study was to determine an optimum pretreatment condition for reducing sugar production, which could be further fermented by yeast to produce ethanol. Preliminary study was conducted to determine the range of duration of heating and temperature that will be used in the optimization by using response surface methodology (RSM). Central composite design (CCD) was used with three independent variables (duration of heating, temperature and acid concentration). Reducing sugar yield per initial biomass was used as a response variable. Preliminary study, that was conducted at 160, 170, 180, 190, and 200 °C for 5, 7.5, 10, 12.5, and 15 min, shows that pretreatment at temperature of 170–190 °C for 5–10 min produced higher reducing sugars than other conditions. Optimization using RSM shows that the optimum condition of MOA pretreatment of OPEFB was at 190 °C for 3 min with 1.1% oxalic acid, which resulted in as much as 34.60% reducing sugars after enzymatic saccharification. The pretreated OPEFB was then characterized and compared with untreated OPEFB. MOA pretreatment successfully removed 50.57% of lignin and 76.56% of hemicellulose from the OPEFB that were confirmed by a decrease or disappearance of the absorption bands of functional groups at 1339–1650 cm−1 and 1735 cm−1, respectively.

The biodegradation and biodetoxification of batik industrial wastewater by laccase enzyme immobilised on light expanded clay aggregate (LECA) were investigated. Laccase from Trametes hirsuta EDN 082 was covalently immobilised by modifying the LECA surface using (3–aminopropyl)trimethoxysilane and glutaraldehyde. The enzymatic characterisation of LECA–laccase showed promising results with an enzyme loading of 6.67 U/g and an immobilisation yield of 66.7% at the initial laccase activity of 10 U/g LECA. LECA-laccase successfully degraded batik industrial wastewater containing indigosol dye up to 98.2%. In addition, the decolorisation extent was more than 95.4% after four cycles. The phytotoxicity assessment of Vigna radiata and the microbial toxicity of two pathogenic bacteria, Bacillus subtilis and Pseudomonas aeruginosa, showed biodetoxification of treated batik dye wastewater. The characterisation using 3D light microscopy, scanning electron microscopy and Fourier transform infrared for LECA–laccase confirmed that laccase was successfully immobilised on LECA, and the decolorisation achieved through the combination of adsorption and enzymatic degradation. This study offers an environmentally friendly, effective and affordable LECA–laccase as a method for batik dye wastewater treatment. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02806-8.

PURPOSE: The purpose of this study was to develop antimicrobial bioplastics based on Poly Lactic Acid (PLA) with the addition of chitosan-ZnO, and chitosan-TiO(2) to improve antimicrobial properties. METHODS: For the preparation of the bioplastics, PLA with chitosan-ZnO or chitosan-TiO(2) were used. The antimicrobial activity, mechanical and thermal properties, and water vapor permeability of bioplastics were evaluated. RESULTS: PLA-chitosan-ZnO indicated a robust antimicrobial activity against bacteria such as Salmonella typhi, Bacillus subtilis, Escherichia coli, Staphylococcus aureus, yeast such as Candida albicans, and fungus Aspergillus niger. No formation of new functional groups in PLA-chitosan-ZnO composites. In comparison to other PLA-based bioplastics, this bioplastic has medium tensile strength, tensile modulus, and elongation percentages with low barrier ability to water vapor. Chitosan-ZnO itself has a greater tensile strength compared to chitosan-TiO(2). These two compounds undergo 2 stages of decomposition in a temperature range of 43 °C to 265 °C. The addition of PLA into chitosan-ZnO or chitosan TiO(2) causes the bioplastics decomposed in a single stage. It also increases the decomposition temperature of bioplastic. However, compared to chitosan-ZnO or TiO(2), the PLA-chitosan-ZnO or TiO(2) bioplastics tend to produce a fragile composite indicating by decrease in their tensile strength. CONCLUSION: In general, the addition of chitosan-ZnO into in PLA-based bioplastic produces better antimicrobial properties compared to TiO(2).

Limbah pewarna sintetik yang dihasilkan oleh industri tekstil dapat membahayakan ekosistem perairan dan organismenya pada konsentrasi tertentu. Proses dekolorisasi secara biologis menjadi fokus penelitian dalam beberapa tahun terakhir karena prosesnya yang relatif murah dan ramah lingkungan. Cendawan pelapuk putih merupakan salah satu mikroorganisme yang dimanfaatkan untuk memproses limbah pewarna melalui proses enzimatik maupun adsorpsi. Fokus dari review ini adalah untuk membahas cendawan pelapuk putih asal indonesia yang berpotensi sebagai agen biodekolorisasi limbah pewarna industri tekstil. Beberapa spesies cendawan pelapuk putih asal indonesia dari genus Ganoderma, Trametes, Pleurotus, dan Leiotrametes telah dilaporkan memiliki kemampuan mendekolorisasi beberapa jenis pewarna sintetik golongan antrakuinon dan azo. Faktor-faktor seperti jenis cendawan, pH, mediator, aktivitas dan jenis enzim, konsentrasi dan jenis pewarna yang diujikan, waktu inkubasi, dan teknik imobilisasi miselia atau ekstrak enzim ligninolitik berpengaruh terhadap proses dekolorisasi secara biologis.

The coating process on food packaging paper is carried out to improve the food safety aspect from dangerous substance migration from food packaging into food. It can facilitate to increase its function and wide the potency of utilization. The coating materials commonly used are plastic and wax coatings. Chitosan as a natural polymer that has the binding properties both water and oil. Chitosan has polar and non-polar groups thus it is potential to be developed as a coating material for food packaging paper. The purpose of this study was to determine the physical-mechanical and antimicrobial properties of food packaging paper from oil palm empty fruit bunches pulp coated with chitosan. The parameters used were chitosan and pulp weight. The oil palm empty fruit bunch pulp was fibrillated with high speed blender then mixed with chitosan solution. The mixed solution was poured in the mold and dried in an oven at 60 °C for 24 hours. The results showed that increasing of chitosan content was coated paper increased physical and mechanical properties values. The more pulp content the higher the physical properties value, however the mechanical properties value tend to decrease. The antimicrobial activity of chitosan coated paper was evaluated by measuring the diameter of the inhibition zones. The results revealed that all tested samples showed growth inhibition zone againts to Escherichia coli and Staphylococcus aureus after 24 h incubation compared to commercial food packaging paper.

Pretreated of sugarcane bagasse was used as a carrier for immobilization of Saccharomyces cerevisiae. Pretreatments were carried out by steaming, pressurized steam, and combination both of procedure. The objectives of this research was to investigate the effect of pretreatment on sugarcane bagasse to cells adsorption and bioethanol production. Immobilization process was conducted in a ratio of 2.5 g carrier/50 mL cell suspension. Whole cell biocatalyst as much as 1% (w/v) was used as inoculum for bioethanol fermentation. The best pretreated sugarcane bagasse for carrier of immobilized cells was obtained using steam treatment for 30 minutes. Those treatment improved the physical properties of carrier and increased the cell retention up to 10.05 mg/g. The use of whole cell biocatalyst after steaming pretreatment also enhanced ethanol yield 1.5 times higher than control. Keywords: bioethanol; immobilization; pretreatment; steam treatment; sugarcane bagasse

The ability of ligninolytic enzymes from Trametes hirsuta D7 to decolorize several synthetic dyes was investigated. A crude enzyme powder was produced by fermenting oil palm empty fruit bunch fibers for one month. The dye decolorization by the enzymes occurred at an efficient 0.25 U/mL. The enzymes degraded 100, 200, 300, 400, and 500 ppm Remazol Brilliant Blue R (RBBR) within 7 h by 95.57 ± 0.32%, 93.46 ± 3.09%, 91.84 ± 0.65%, 86.44 ± 0.97%, and 82.14 ± 0.52%, respectively. The enzyme also decolorized anthraquinone (Acid Blue 129), monoazo (Acid Orange 7), diazo (Reactive Black 5), and trimethyl methane (Methyl Violet) dyes within 7 h by 94.59 ± 7.97%, 13.99 ± 0.30%, 7.61 ± 0.01%, and 7.59 ± 0.18%, respectively. Addition of MnSO 4 , H 2 O 2 , and violuric acid enhanced the dye decolorization rate by up to 10-fold. This study shows the potential for application of ligninolytic enzymes from T. hirsuta D7 in the treatment of wastewater effluent of textile industries.

Biorefinery concept encourages agricultural sector for developing zero waste technology. Utilization of agricultural residues for producing valuable products has become an important strategy to be investigated. Sorghum (Sorghum bicolor L) biomass, as one of agricultural residues, is potentially of great significance as a lignocellulosic material for production of lignocellulolytic enzymessuch as Laccase (Lac), Manganese Peroxidase (MnP), and Lignin Peroxidase (LiP), Cellulases (Cel), and Xylanases (Xyl). In the present study, 13 Indonesian sorghum accessions (with different lignocellulosic characteristics) were investigated due to their potency as potential substrate for production ligninolytic, hemicellulolytic, and cellulolytic enzymes by Trametes hirsuta AA-017 which has ability to sequentially produce the enzymes. Three accessions (Samurai, 4183, Kawali) were found as most suitable materials for enzymes production by the fungus with highest enhancement were more than 8000 and 71-fold, respectively for Lac and MnP. Total lignin in the biomass has shown a positive correlation with Lac (Pearson coefficient = 0.630; P

The influence of sulfuric acid concentration in aqueous glycerol during microwave pretreatment on the chemical content and morphological characteristics changes of pretreated EFB were investigated. After pretreatment, the pulp was further subjected to enzymatic hydrolysis. The micro-morphology and crystallinity of original and pretreated EFB were observed by scanning electron microscopy (SEM) and X-ray diffraction. Lignin content of pretreated EFB was determined, while hydrolysate after enzymatic hydrolysis was also analyzed the reducing sugar content. These results indicated that microwave assisted acid pretreatment of EFB caused weight loss. Evaluation on effect of varying acid concentration and residence times results 1 % acid concentration for 12.5 min as the best pretreatment condition. Under the best pretreatment condition, lignin removal was about 59.16 % with the reducing sugar yield per original EFB was 25.09 or 55.85 % per pretreated EFB. The acid–glycerol catalyst successfully improved reducing sugar yield compared to both untreated and biological pretreated samples. SEM images revealed that the pretreated of EFB surface changed noticeably and the increase of crystallinity degree of pretreated EFB indicated a great effect on amorphous region to crystalline region. The addition glycerol in reaction mixture both acid and alkaline has synergistic effect to increase reducing sugar yield. Much volume of glycerol in mixture has great effect to improve enzymatic hydrolysis performance than that of acid ones. The highest reducing sugar yield is produced as much as 98 % when EFB irradiated using glycerol–acid ratio of 1:2 for 12.5 min at 1 % acid concentration. The addition glycerol of 2 fold can increase 3.9 fold reducing sugar yield in the best pretreatment condition. Microwave assisted acid pretreatment in aqueous glycerol was an effective way to increase reducing sugar yield of EFB by lignin removal and fiber disruption.

This research was intended to study the effect of Amphiphilic Lignin Derivatives (A-LD) surfactant addition on the bioethanol production by using simultaneous saccharification and fermentation (SSF) system. A-LD synthesis was obtained from single-step lignin isolation from black liquor as a by-product of the Acacia mangium kraft process. This synthesis was carried out by reacting single-step lignin isolation with the polyethylene glycoldiglycidylether (PEDGE), which was heated at 60 °C for one h. SSF was performed by using Saccharomyces cerevisiae InaCC Y93, cellulase enzyme of 50 FPU/g, yeast inoculum in 10% and 20%, and a 15 mg/mL A-LD added into 5.37 g (wet weight) of sweet sorghum bagasse (SSB) kraft pulp. The concentration of reducing sugar and the ethanol contents were analyzed every 24 h for 72 h and also at 89 h. The ethanol concentration, ethanol yield, and the highest percentage of theoretical ethanol yield were 4.91 ± 0.10 g/L, 0.06 ± 0.001 g/g, and 11.29 ± 0.22%, respectively. They were obtained by 20% of yeast inoculum for 24 h fermentation on the SSF system and the addition of A-LD surfactant. The addition of A-LD on the SSF process increased ethanol production and shortened the fermentation time.

Laccase is one important enzyme in decolorization of textile dyes. This research aims to study the optimization of laccase production from Trametes sp. EDN134 under several variation of substrates (bagasse, bamboo, Jabon wood, oil palm empty fruit bunch (OPEFB), and sorghum fibers), time incubation (1-15 d), growth temperature (25, 27, 30, 32, 35 and 37°C), and concentrations of inducer CuSO4 (1, 2, 3, 4 and 5 mM). The results showed that optimum laccase specific activity (14,838 U/g) detected in the OPEFB medium at 10 days incubation, temperature 30°C and CuSO4 2 mM. The study suggests that these parameters could be used for the optimization of laccase production from a newly isolated Trametes sp. EDN134.

EFFECTS OF CHITOSAN COATING ON THE PHYSICAL, MECHANICAL AND ANTIMICROBIAL PROPERTIES OF FOOD PACKAGING PAPER. The coating process on food packaging paper is carried out to improve the food safety and health aspect from dangerous substance migration from food packaging into food. Chitosan has attracted interest in packaging, especially in food packaging as edible films and coatings. A paper from oil palm empty fruit bunches (OPEFB) pulp was coated with chitosan using different pulp weight (1, 1.5, 2 g) and chitosan content (0.25, 0.50, 0.75, 1 g) as parameters. The effect of chitosan as coating material on physical, mechanical, and antimicrobial properties was studied. The results showed that the density and grammage values of the paper increased after coating due to the increasing of chitosan content. The mechanical properties of the coated paper, such as tensile strength, tensile modulus and elongation, improved in line with the increase of chitosan content and pulp weight. The addition of chitosan on paper imparts antimicrobial properties against Gram-positive bacteria (Staphylococcus aureus) and Gram- negative bacteria (Escherichia coli).

Penggunaan kultur campuran jamur pelapuk putih pada proses pretreatment bahan lignoselulosa belum banyak diteliti. Penelitian ini bertujuan untuk mempelajari pengaruh penggunaan kultur Phanerochaete chrysosporium dan Trametes versicolor pada proses pretreatment serat tandan kosong dan pelepah kelapa sawit terhadap hasil sakarifikasinya. Inokulum P. Chrysosporium dan T. versicolor dituangkan ke dalam sampel substrat serat tandan kosong dan pelepah kelapa sawit (40-60 mesh) yang telah disterilkan, masing-masing sebanyak 5% (w/v), sehingga total inokulum yang ditambahkan ke dalam kedua macam substrat masing-masing 10% (w/v). Sampel diinkubasikan pada suhu ±27°C selama 4 minggu. Sebagian dari contoh diambil dan lalu diperiksa setelah masa inkubasi 1, 2, 3, dan 4 minggu. Sakarifikasi dilakukan menggunakan enzim selulase sebanyak 20 FPU per g biomassa dan β-glukosidase dalam shaking waterbath pada suhu 50°C selama 48 jam. Analisis gula pereduksi, glukosa dan xilosa dilakukan terhadap hasil sakarifikasi. Rendemen gula pereduksi, konsentrasi glukosa dan xilosa tertinggi dari tandan kosong kelapa sawit. diperoleh dari sakarifikasi serat dengan pretreatment selama 4 minggu, yaitu masing-masing 13,08%, 0,86 mg/g dan 0,13 mg/g, sedangkan rendemen gula pereduksi, konsentrasi glukosa dan xilosa tertinggi dari pelepah kelapa sawit didapatkan dari sakarifikasi substrat dengan pretreatment selama 2 minggu, yaitu masing-masing 8,98%, 0,92 mg/g dan 0,23 mg/g.

Decolorization of synthetic dyes by laccase produced from newly isolated Trametes hirsuta EDN084 under in-vitro condition was investigated in this study. Partial purification was conducted using ultrafiltration Amicon 10K at 5000 x g for 20 min. Laccase (0.1 U/mL) could decolorize 50% remazol brilliant blue R (RBBR), 47% reactive blue 4 (RBlue4), 51% acid blue 129 (AB129), 21% acid blue 25 (AB25), 40% acid blue 113 (AB113), 11% acid orange 7 (AO7), 2% reactive black 5 (RBlack5), 2% reactive red 120 (RR120), and 85% direct blue 71 (DBlue71) for 4 h. In order to improve the decolorization, the addition of violuric acid (VA), 2, 2, 6, 6-tetramethylpiperidine 1-oxyl (TEMPO), and 1-hydroxybenzotriazole (HBT) were individually assayed. The result showed that decolorization was improved significantly (2 to 30-fold) after the addition of 1 mM VA. This study suggests that mediator VA is suitable for the enhanced decolorization of synthetic dyes by laccase from T. hirsuta EDN084.

Light Expanded Clay Aggregate (LECA) in a granular form was used for the immobilization of fungus Trametes hirsuta D7 to decolorize Remazol Brilliant Blue R (RBBR) dye. The efficacy of LECA as a media for immobilization was assessed using steam activation, contact time, immobilization method and dosage of sorbent. Batch experiments were carried out for decolorization and the results showed that activated LECA has the capability to remove 35% RBBR for 24 h of contact time. Non-freeze dry immobilization process demonstrated a higher decolorization than that of freeze dry process with maximum 88% and 22% removal, respectively. Different dosage of activated and non-freeze dry-LECA was tested to achieve optimum decolorization and it was found that 0.2 g/ml dye could remove 76% dye for 3 hours and reached the maximum removal for 94% at 24 hours with high laccase activities (460 U L− 1). This study found that fungus T. hirsuta D7 and LECA could be used to decolorize RBBR and LECA had the potential to be an alternative matrix for fungi immobilization.

Styrofoam wastes are composed of many polymerized styrene monomers that are generally considered to be recalcitrant and are resistant to biodegradation. In this study, the ability of ligninolytic fungi and bacteria were investigated on degradation of styrofoam wastes. All the fungi and bacteria used were able to grow on agar media containing styrofoam. Fungi Cymatoderma dendriticum WM01, Ceriporia sp. BIOM3, and Pestalotiopsis sp. NG007 degraded 15.7%, 19.4%, and 74.4% styrofoam within 30 d, respectively. Cerratia marcescens BLSP4, Bacillus subtilis BLSP4, and Pseudomonas aeruginosa BLSP4 degraded 38.3%, 52.6%, and 63.4% styrofoam, respectively. SEM analysis demonstrated the appearance of micro pore in styrofoams treated with Pestalotiopsis sp. NG007 and P. aeruginosa indicating biodegradation. In addition, analysis using UATR FTIR corroborated removal of some functional groups from the degraded styrofoam were eliminated. This study showed strains of ligninolytic fungi and selected bacteria have the potential to be used in bioremediation of styrofoam wastes.

Previous research on anatomical structures of pretreated large (betung) bamboo (Dendrocalamusasper) using single culture of white-rot fungi has been investigated, which revealed that the pretreatment caused the decrease in the Runkel ratioas well as the coefficient rigidity and the increase in the flexibility ratio of their corresponding bamboo fibers. However, there is no study reported on the anatomical structure changes of them caused by pretreatment using mixed culture of white-rot fungi. This paper reports the results of the research on paper/pulp quality after different treatments. Pretreatment that used Trametes versicolor fungi and lasted for 45 days inflicted intensive fiber damages compared with those of untreated bamboo (control). Fresh and barkless large (betung) bamboo chips of 2 year's old, and 1.6 cm in length, were inoculated by 10% of mixed culture of white-rot fungi inoculums stock for 30 and 45 days in room temperature. There were four treatment groups of mixed culture, i.e T. versi color and P. ostreatus (TVPO); P. ostreatus and P. chrysosporium (POPC); P. chrysosporium and T.versi color (PCTV); and P.chrysosporium, T.versicolorand P.ostreatus (TVPCPO).After the inoculation period, the chips weremacerated into separate fibers using Scultze method to analyze the fiber dimension and its derived values. The fibers were then observed regarding their macro and microscopic structures by optical microscope. Mixed culture pretreatment of white-rot fungi accelerated improvement of fiber morphology and fiber derived value characteristics, except for Muhlsteph ratio. The fiber derived values oftreated bamboo tended to improve compared to those of untreated bamboo, there by requiring milder pulping conditions. Accordingly, the treated bamboo would indicatively produce a good quality pulp (grade I) based on FAO and LPHH (Forest Product Research Report) requirements. Co-culture treatment using P. chrysosporium and P. ostreatus for 45 days produced the best fiber dimension and its derived value properties. The fungi hypae colonized on the surface area of bamboo followed by mycelium penetration into substrate (bamboo-inner structure). The partial degradation caused by delignification indicatively attributed to the fungi activity was shown in the macroscopic images.

Conversion of lignocellulosic biomass requires pretreatment in order to separate lignin from cellulose. Several methods have been introduced for pretreatment process of lignocellulosic biomass. These are physical, physico-chemical, chemical, as well as biological pretreatment. The following investigation aimed at a biological pretreatment of bagasse using mix-culture of white-rot fungi with the objective to investigate pretreatment effect on the lignin degradation of bagasse with minimum loss of cellulose content. The white-rot fungi involved Phanerochaete crysosporium (Pc), Pleurotus ostreatus (Po), and Trametes versicolor (Tv) on degrading lignin of bagasse with minimal losses of cellulose content 5% (w/v). The culture variation are as follows Po dan Tv (1:1), Po dan Pc (1:1), Pc dan Tv (1:1) and Po, Pc dan Tv (1:1:1) they were inoculated into steamed bagasse and incubated for 1, 2, 3, and 4 weeks respectively. Chemical analysis was done to determine the weight loss, lignin, and alphacellulose content before and after pretreatment. Optimum lignin degradation was achieved after two weeks incubation for each mix-culture. The most effective lignin degradation of mix-culture Po, Pc and Tv (25,26±1,57%) with minimum cellulose loss (17,27±3,76%). Key words : pretreatment, white-rot fungi, mix-culture, baggase INTISARI Konversi biomassa lignoselulosa membutuhkan proses perlakuan pendahuluan yang bertujuan untuk memisahkan lignin dari selulosa. Beberapa metode telah banyak diketahui untuk proses perlakuan pendahuluan biomassa lignoselulosa, diantaranya perlakuan pendahuluan secara fisika, fisika-kimia, kimia dan biologi. Penelitian ini bertujuan untuk mengetahui pengaruh penggunaan kultur campur jamur pelapuk putih Phanerochaete crysosporium (Pc), Pleurotus ostreatus (Po), dan Trametes versicolor (Tv) terhadap penurunan kadar lignin bagas dengan kehilangan minimal α-selulosa. Inokulum yang digunakan sebesar 5% (w/v) dengan variasi kultur Po dan Tv (1:1), Po dan Pc (1:1), Pc dan Tv (1:1) dan Po, Pc dan Tv (1:1:1). Kultur campur jamur diinokulasikan ke dalam bagas dan diinkubasi masing-masing selama 1, 2, 3, dan 4 minggu. Analisa kimia dilakukan untuk mengetahui kehilangan berat, kadar lignin serta selulosa bagas sebelum dan setelah perlakuan pendahuluan. Hasil penelitian menunjukkan bahwa penurunan optimal kadar lignin bagas dengan kehilangan minimal α-selulosa terjadi pada waktu inkubasi dua minggu untuk setiap variasi kultur campur. Dari empat variasi kultur campur pada waktu inkubasi dua minggu menunjukkan bahwa kehilangan rata-rata lignin bagas tertinggi (325,26±1,57%) dengan kehilangan minimal α-selulosa (17,27±3,76%) terjadi pada kultur campur Pc dan Tv. Kata kunci : perlakuan pendahuluan, jamur pelapuk putih, kultur campur, bagas

Lindi hitam merupakan limbah industri pulp yang belum termanfaatkan dengan baik. Pemanfaatan lignin dari lindi hitam selama ini biasanya hanya digunakan sebagai perekat. Dalam bidang mikrobiologi lignin dari lindi hitam dapat dimanfaatkan sebagai media selektif untuk isolasi jamur pelapuk putih. Tujuan penelitian ini adalah untuk mengetahui manfaat lignin hasil isolasi proses biopulping sebagai media selektif untuk jamur pelapuk putih. Lignin dari lindi hitam hasil biopulping bambu, dengan proses pemasakan soda dan kraft, diisolasi dengan penambahan asam. Padatan lignin kemudian dimurnikan menggunakan larutan dioksan dan ditimbang berat serta dianalisa secara kualitatif menggunakan spektrofotometer. Lignin hasil isolasi ditambahkan pada media agar untuk uji selektifitas jamur pelapuk putih Phanerochaete crysosporium dan Trametes versicolor. Pretreatment bambu pada 30 hari inkubasi menghasilkan lignin yang lebih tinggi jika dibandingkan dengan inkubasi selama 45 hari. Padatan lignin yang diperoleh dari hasil biopulping proses kraft juga lebih tinggi jika dibandingkan pada proses soda. Pengujian selektifitas jamur pada media alkali lignin menunjukkan bahwa fungi T. versicolor mensekresi enzim lebih cepat daripada P. chrysosporium.

ENHANCEMENT OF REDUCING SUGAR PRODUCTION FROM OIL PALM EMPTY FRUIT BUNCH BY PRETREATMENT USING ORGANIC ACID IN PRESSURIZED REACTOR . Organic acids are potential to create more environmentally friendly process in the pretreatment of lignocellulosic biomass for bioethanol production. This study was aimed to investigate the influence of organic acid pretreatment in reducing sugar production in a pressurized reactor with various resident times and temperatures on enzymatic hydrolysis of OPEFB. Two different organic acids (maleic acid and oxalic acid) were used in the pretreatment of oil palm empty fruit bunch (OPEFB) using a pressurized reactor. Factorial design using three different temperatures (1 7 0, 1 8 0, and 1 9 0°C) and four resident times ( 15 , 30 , 45 , and 60 min) were employed, followed by enzymatic hydrolysis. Each condition conducted two repetitions. Analysis was conducted on the reducing sugar that was produced after saccharification by means of the severity factor of each pretreatment condition. Maleic acid showed higher reducing sugar yield with lower severity factor than oxalic acid with the same operating conditions. The highest yield of reducing sugars (80.84%) was obtained using maleic acid at 170 for 60 minutes with severity factor of 1.836. Keywords: bioethanol; organic acid pretreatment; pressurized reactor; severity factor; oil palm empty fruit bunches; Abstrak Asam organik berpotensi dalam membantu proses praperlakuan dari biomassa lignoselulosa untuk memproduksi bioetanol yang ramah lingkungan. Penelitian ini bertujuan untuk mengetahui pengaruh asam organik, suhu dan waktu operasi terhadap produksi gula pereduksi dengan reaktor bertekanan pada tandan kosong kelapa sawit. Dua asam organik yang berbeda yaitu asam oksalat dan asam maleat digunakan untuk proses praperlakuan tandan kosong kelapa sawit (TKKS) dengan bantuan reaktor bertekanan. Dalam proses praperlakuan digunakan tiga suhu yang berbeda yaitu suhu 170, 180, dan 190°C dan empat waktu operasi 15, 30, 45, dan 60 min yang dilanjutkan dengan proses hidrolisis enzimatis. Setiap kondisi dilakukan dua kali pengulangan. Analisa yang digunakan adalah analisa uji gula pereduksi dan severity factor pada kondisi tiap praperlakuan. Asam maleat menunjukkan hasil yang lebih baik dengan severity factor yang lebih rendah dibandingkan menggunakan asam oksalat dengan kondisi operasi yang sama. Hasil yang didapatkan menunjukkan bahwa praperlakuan tandan kosong kelapa sawit dengan bantuan reaktor bertekanan memiliki rendemen gula pereduksi optimum sebesar 80,84% dengan menggunakan asam maleat pada suhu 170 ° C selama 60 menit dengan severity factor sebesar 1,836. Kata kunci: bioetanol; praperlakuan asam organik; reaktor bertekanan; severity factor; tandan kosong kelapa sawit.

Oil palm frond is one type of lignocellulosic biomass abundantly and daily available in Indonesia. It contains cellulose which can be converted to glucose, and further processed to produce different kinds of value –added products. The aim of this research is to study the effects of biological pretreatment of oil palm frond (OPF) fiber using Phanerochaete chrysosporium and Trametes versicolor on the enzymatic saccharification of the biomass. The OPF fiber (40-60 mesh sizes) was inoculated with cultures of the two fungi and incubated at 27 °C for 4 weeks. The samples were taken after 1, 2, 3, and 4 weeks of incubation. Chemical components of the biomass after pretreatment were analyzed. The saccharification of the pretreated samples using cellulase and β-glucosidase was performed in a water bath shaker at 50 °C for 48 hours. The concentration of reducing sugar increased with increasing of incubation time, either in those pretreated with culture of P. chrysosporium or with T. versicolor. Pretreatment of OPF fiber using single culture of T. versicolor for 4 weeks gave the highest reducing sugar yield (12.61% of dry biomass).

Sugarcane bagasse was used as a carrier to immobilize Saccharomyces cerevisiae in bioethanol production. This research aims to study the potential use of sugarcane bagasse as an alternative carrier for cell immobilization and improvement in the production process of cell immobilization in bagasse. The results showed that the physical characteristics of sugarcane bagasse as a carrier were water content (7.77 ± 0.35%), water retention (4.80 ± 0.44 g/g), water absorption index (8.58 ± 0.22 g/g), and lignin content (24.40 ± 1.52 %). Determination of cell retention was performed in an inoculum volume of 50 mL yeast suspension with various carrier weights (2.5, 5, 10, and 20 g). The highest cell retention was obtained in ratio of 2.5g carrier/50 mL cell suspension with cell retention of 5.41 ±1.06 mg/g, or known as biocatalyst. Biocatalyst, as much as 1.5, 3, 4.5, and 6 g, were used as inoculum for a 24 hour bioethanol fermentation. The best concentration and productivity of bioethanol, obtained by using 3 g of biocatalyst, were 23.95 ±0.28 g/L and 1.24 ±0.01 g/L/hours. The average of bioethanol yield for a 24 hour fermentation by using immobilized cells was three times higher than the free cells system.