Production of Xylitol From Sago Lignocellulosic Waste

Destarched sago fibre which contained lignocellulosic material can be exploited for the production of value-added product such as xylitol. Microbial xylitol production has received much attention because of its flexibility in term of adaptation, ease of operation and eco-friendliness. The aim of this study is to produce xylitol from sago lignocellulosic waste by focusing on the pre-treatment technique and fermentation parameters. Pre-treatment parameters such as dilute sulphuric acid concentration (0% (v/v) - 6% (v/v), solid to liquid ratio of hydrolysis system (5:100 – 40:100), hydrolysis process reaction time (30 – 90 minutes) using high-pressure steam pre-treatment, mild hydrothermal pre-treatment, and microwave pre-treatment were studied. The hydrolysate obtained from the selected pre-treatment was characterised and selected for fermentation. In order to obtained the highest xylitol production, fermentation parameters such as inoculum age (9, 18, 27 hours), inoculum concentration (10 – 30%), and effects of yeast extract and peptone were studied. The result revealed that pre-treatment using high pressure steam produced the highest xylose concentration with acid concentration of 2% v/v, reaction time of 30 minutes and solid to liquid ratio 30:100. The acid hydrolysis which was integrated with high pressure steam pre-treatment exhibits highest xylose (11.56 g/L) and glucose (22.51 g/L) production in which the temperature was set at 121℃ and solid to liquid ratio of 30:100. The leftover hydrolysate from pre-treatment of high-pressure steam were then used for xylitol fermentation using C. tropicalis. The inoculum age of 28 hour with inoculum level of 20% was chosen as it consumes glucose and xylose efficiently in the hydrolysate without any supplementation and produces highest xylitol concentration of 0.86 ± 0.11 g/L compared with the control experiments containing commercial xylose and glucose which produced xylitol concentration of 2.56 ± 0.11 g/L. The fermentation of destarched sago fibre hydrolysate with addition of yeast extract significantly increases the consumption of glucose and xylose and produces maximum xylitol production which was 11.75 ± 0.09 g/L which was higher than the control experiment which produced 7.07 ± 0.06 g/L after 60 h of batch fermentation process. This work indicates that the optimal pre-treatment of destarched sago fibre and selected parameter of fermentation of C. tropicalis can influences xylitol production. The studies suggests that destarched sago fibre can be potentially served as an alternative raw material for xylitol production

Digital Comics as an Alternative Teaching and Assessment Tool for Biotechnology Courses

Storytelling has been long applied in teaching and learning. Nevertheless, application of storytelling through digital comics for teaching and learning Biotechnology is still limitedly explored. This presentation highlights the initiative of using digital comics as an alternative teaching and assessment tool for Bioprocess Technology and Environmental Biotechnology. The courses were taken by 136 students of Resource Biotechnology programme at Faculty of Resource Science and Technology, UNIMAS in Semester 1 and 2 2021/2022. Engaging digital comics entitled Ahmad’s FYP story, To RIP: Landfills or Composting?, Save Our Souls and From Textile Waste to Biofuel were topics of Environmental Biotechnology. The resulting comics produced by the students were disseminated to selected schools and matriculation centres and also in social media. Upon the created and used in the teaching and learning session of the aforementioned courses. Questions that assess students’ understanding were also integrated in the comics in order to engage the students to think and reflect whilst reading the comics. We have also used comics as a means of a graded assessment whereby the students were required to produce digital comics related to the implementation of the approach, the students’ average scores for both courses have improved by 30 to 36%. This is further supported by the positive feedback from the students indicating good acceptance of the approach. The initiative has also resulted in several added values amongst the students such as enhancement of art skill, digital skill, science communication skill, storytelling skill, community responsibility besides promoting the culture of learning during leisure time. Moreover, the comics also have a potential to be used as life-long learning materials in both formal and informal contexts besides having the commercial potential to be marketed in the form of digital or printed comics. In summary, the positive impacts of this initiative have clearly proved the efficacy of digital comics as an alternative teaching and assessment tool for Biotechnology courses

Production of High Quality Silage from Sago Fronds

Sago frond is one of the most abundant by-products from the sago industry. Sago fronds are left on the ground upon harvesting of the sago palm for production of sago flour. Over 500 palms are harvested every day in the district of Mukah, Sarawak to feed the numerous sago mills. From here, it has been approximated that over 26 tons of sago frond is discarded as a rotting biomass. We see the potentials of sago frond to be developed as the alternative material for large-scale production of lactic acid and as a feed material for the livestock industries. Sago frond contains sap that can be extracted from the de-skinned white pith using a roller compressor, akin to sugar cane press. The sap contains 7% free sugars consist mainly of glucose and xylose. Both sugars are ideal substrates - and have been used - for the production of lactic acid utilising lactic acid bacteria (LAB) such as Lactococcus lactis. The residual fibre can be used as high-quality animal feed with minimal treatment. However, feed quality and palatability is very much improved through lactate fermentation during ensiling to produce silage. Ensiling process require anaerobic condition to inhibit the growth of moulds and fungi that can spoil the silage while concomitantly allowing the growth of lactic acid bacteria to produce nutrient and increase digestibility of the silage. The ensiling process takes only three weeks, whereby the pH of the silage is reduced to 4.4, a definitive condition that inhibits the growth of mould and fungi. Thus, addition of antibiotic and antifungal which usually entails the usual production of animal feed is unnecessary. This ensures that sago frond silage is much healthier and safer for consumption by the livestock. Silage from sago frond contains 9% carbohydrate and 18% protein, a balanced ratio of carbohydrate and protein highly recommended for growing and lactating stages of goats. Ensiling process of sago frond can be improved by using the LAB cells from the production of lactic acid on sago frond sap to boost the ensiling process. Adding LAB cells will hasten acidification to lower the pH, which will reduce duration of the ensiling process together with augmenting its digestibility. These findings are crucial in maximising the use of sago fronds – either disposed upon daily trimming or perpetual harvesting of matured logs – which proves to be a sustainable source for both the animal feed and lactate industries. With these possibilities, meagre income of the sago farmers can be enhanced, followed by development of new sago plantations which focus on zero wastes to protect our environment

Effect of medium supplementation on very high gravity bioethanol fermentation using sago hampas hydrolysate as a feedstock

Bioethanol fermentation under very high gravity (VHG) conditions by using sago hampas hydrolysate (SHH) supplemented with yeast extract as a fermentation medium has resulted in an incomplete metabolism of glucose, leading to relatively low bioethanol production in comparison to the theoretical titre. Therefore, additional supplementation of the fermentation medium is necessary to increase the yeast tolerance towards inhibitors and high concentration of glucose and bioethanol. This work investigates the effect of supplementing SHH media with various nutrients on bioethanol fermentation under VHG conditions. The nutrients included magnesium sulphate (0.12 g/L), urea (3 g/L), glutamic acid (5 g/L), and peptone (5 g/L). Our results showed that culture supplemented with peptone has significantly improved the yeast growth by 0.9-fold and glucose consumption efficiency by 10% compared to the control cultures. Besides that, the media formulation has also increased bioethanol production by 13%, with a maximum concentration of 126.20 ± 3.0 g/L. In general, the results suggest an improved formulation of fermentation medium consisting of SHH for bioethanol production under VHG conditions. These results will provide useful insights into the development of bioethanol production from sago-based feedstock in the future

Feasibility of Pre-Harvest Sago Frond as State-of-the-Art Resources to Produce Animal Feed

Global crises such as climate change, war and borderless diseases are the factors that lead to limited supply and unstable prices of the raw material to produce animal feed placing a major burden on the farmers and smallholders to produce livestock at a reliable cost. Malaysia is among the most affected country due to its high dependency on imported resources to produce animal feed. Hence, cheap and locally available raw material is the key to producing sustainable and safety-ensured animal feed for domestic consumption. Due to the adaptability and resilient nature of sago palm towards extreme environmental distress, sago frond was selected as an alternative raw material to produce animal feed in the form of silage. Sago frond was pruned from growing sago palm (age between 3-7 years), then leaves and rachis were pulverised before vacuum packed into silo bag. Analysis shows that optimised sago frond silage (1:1 RSF/SL) possessed five ideal characteristics that include dry matter (47.76 %), acid detergent fibre (31.98 %), total water-soluble sugar (2.4 g/Kg), minimum pH (4.3) and protein content (16.85 %). Hence, the ensiling technique applies to produce high-quality animal feed from optimum formulated sago frond by preserving nutrient content and improving in-vitro digestibility of the silage designated for ruminant consumption. The feeding trial shows significant growth performance of animal models (Malin Breed Sheep) fed with optimised SFSil with Average Daily Gain (61.12 g/day) and Feed Conversion Ratio (9.64 g/g). Therefore, manufacturing animal feed from pre-harvest sago frond provides the solution to high dependency on imported animal feed and also offers a new lucrative commodity for the sago farmers while waiting for the sago palm to be harvestable

Physicochemical and microbiological assessment of Nypa fruticans sap collected in Sarawak, Malaysia

Nipa sap is a sweet and translucent beverage that originated from nipa palm (Nypa fruticans) tree. In Sarawak, nipa sap become raw material for nipa sugar or locally known as gula apong. However, nipa sap undergoes natural fermentation that alters the properties of nipa sap including taste, aroma and quality. Fermented nipa sap is whitish colour with an unpleasant aroma and taste, which makes it unacceptable for consumption. Hence, it is no longer suitable to make nipa sugar. This study aimed to determine the physicochemical and microbiological changes of nipa palm sap from fresh to fermented. The nipa sap was allowed to undergo natural fermentation at room temperature for 56 days. Samples were collected every 24 hrs for the first week and once a week in the subsequent week. The selected physiochemical qualities were analysed using high-performance liquid chromatography (HPLC) whereas the microbial content was analysed using spread plating. Fresh nipa sap showed the highest load of sugar (334.2±12 g/L) with sucrose as the main sugar found (231.5±4.3 g/L), followed by fructose (42.1±1.2 g/L), and glucose (29.7±3.2 g/L). Fresh nipa sap also possessed the lowest load of ethanol (0.08±0.03 g/L), lactic acid (1.09±0.06 g/L), and acetic acid (0.05±0.01 g/L) as well as microbial and yeast concentration. Later, ethanol started to accumulate on day 4 (9.80±0.1 g/L) and the highest peak was on day 21 (19.1±2.01 g/L). The microbial concentration changed as well, affecting the quality of nipa sap. As nipa sap plays such an important role in the lifestyle of people in Sarawak, this study provides a better understanding of the microbiology and biochemistry of its fermentation process. Hence, proper planning for handling fresh nipa sap should be considered to ensure the quality of value-added product production

THE CHARACTERISTICS OF SAGO FROND SAP FROM TWO SELECTED GROWTH STAGES; ANGKAT PUNGGUNG AND UPONG MUDA PALMS

Sago frond is produced in abundance upon harvesting of the sago palms for starch extraction, hence need to be utilized and developed into beneficial products. In this study, the sap which contains sugars and starch is obtained by roller crushing the skinned frond for use as fermentation medium. Fronds from two selected growth stages (namely Angkat punggung and Upong muda) and two different positions within the rosette (inner and outer circle) of the sago palm were studied. Based on the results, the outer circle frond of Upong muda palm gave the highest volume of sap at 290mL/kg which equivalent to 1600 mL/frond. On top of that, sago frond sap has an acidic pH, with glucose as major sugar component and contained various kinds of minerals like calcium, potassium and manganese. All fronds from two selected growth stages contain glucose between 28-68 g/L and xylose 21-29 g/L, respectively. After 21 days of storage, it can be concluded that the amount of reducing sugars and starch in most samples obtained from two selected growth stages decreased slightly from the original. Subsequently after this study, both fresh and stored sago frond sap can be used as a fermentation substrate without any pre-treatment or modification

Sugars from sago frond as prebiotic substrate to enhance the growth of Lactococcus lactis IO-1 and production of L-lactic acid

Sago palm is often discredited for exhibiting long maturity period and barrenness of pre-harvest products, which restrain its potentials as an alternative and eternal starch provider. The use of sago fronds to produce prebiotic and fermentable sugars from pruned palms and fronds discarded upon harvesting is a possible enterprise to provide income for the cash-strapped sago farmers while waiting for the sago trunks to be harvestable. Dried sago frond powder coupled with the cellulolytic enzyme and incubated for 48 hours, producing a maximum recovery of cellobiose at 25%. This is of great advantage in reducing the cost of large-scale processes since the yield and productivity from SFS is comparable to the Standard Medium and SFS amended with yeast extract at 0.85g/g and 85%, respectively. Meanwhile, the composition of cellobiose as main sugar component increase the viability of the Lactococcus lactis I0- 1 by prolong the lifespan of the cell by perform as slow release carbon source, in fact, cellobiose was protected by β (1-4) glycosidic bond made it consumable to specific probiotic in human digestive system conceive that cellobiose as potential prebiotic component for human. Clearly, the use of sago frond is highly economical and sustainable as the raw material for the manufacturing of fermentable sugars and subsequently as the sustainable substrate for large-scale production of L-lactic acid

Maximising production of prebiotic sugar (Cellobiose) from sago frond

Numerous fronds are discarded as waste upon harvesting of sago logs for starch production. Currently, these fronds are left to degrade in sago estates which potentially pose fire hazard in the dry season, concomitantly accommodating various pests that endanger the livelihood of the sago farmers. The objective of this study is to utilize the frond for the production of cellobiose, a non-table sugar known to harbour various prebiotic properties. Enzymatic hydrolysis was performed on treated sago frond fibre utilizing the cellulolytic enzyme Celluclast 1.5L. Characterization of the lignocellulosic component revealed that adolescent sago fronds have the highest cellulose content (41.43%) which is beneficial for high yield of cellobiose. Pruned sago fronds have the highest lignin (40.63%) which hinders the hydrolysis process. Nevertheless the hemicellulose content was found to be approximately similar (between 15 to18%) which promotes the production of cellobiose. Optimum enzymatic hydrolysis was achieved at 6% (w/v) sago frond powder coupled with 10% (v/v) enzyme and incubated for 48 hours, producing a maximum recovery of cellobiose at 25.5%

Antibacterial properties of purified sago frond sugar against food-borne associated disease bacteria

Sago palm is recognised as key to sustainable food security due to its advantages resilient against extreme conditions such as wildfire and flood associated with adaptability to climate change. Sago palm is also known to remain solid after being attacked by pests and infected by the disease. Unfortunately, for the last ten years, the Sago palm industry experiences a significant decrease in plantation area and productivity. The long maturation period is identified to be the major factor that is responsible towards the respected issue. Thus, alternative commodities from the growing sago palm must be explored to offer a better perspective on the sago industry. Sago frond (SF) was utilised into Sago Frond Sugar (SFS) via enzymatic hydrolysis using cellulase enzyme containing cellobiose and glucose as main sugar at 9-10 g/L and 5-6 g/L concentration respectively. SFS was purified (PSFS) using Powdered Activated Charcoal (PAC) to remove the impurities. Antibacterial analysis shows that PSFS able to inhibit the growth of Staphylococcus aureus, Escherichia coli and Salmonella typhi at 23.5 mm, 22.5mm and 13.25 mm clearing zone respectively. However, the growth of Listeria monocytogenes seems unaffected by the presence of PSFS. Promoting the versatility of sago frond as raw material to synthesise high-value products such as SFS will extend the potential of the sago palm to be recognised as an important crop to ensure global food security and safety