The oil palm sector is one of the major plantation industries in Malaysia. Palm kernel cake is a byproduct of extracted palm kernel oil. Mostly palm kernel cake is wasted or is mixed with other nutrients and used as animal feed, especially for ruminant animals. Recently, palm kernel cake has been identified as an important ingredient for the formulation of animal feed, and it is also exported especially to Europe, South Korea, and Japan. It can barely be consumed by nonruminant (monogastric) animals owing to the high percentages of hemicellulose and cellulose contents. Palm kernel cake must undergo suitable pretreatment in order to decrease the percentage of hemicellulose and cellulose. One of the methods employed in this study is fermentation with microorganisms, particularly fungi, to partially degrade the hemicellulose and cellulose content. This work focused on the production of enzymes by Aspergillus niger and profiling using palm kernel cake as carbon source.
This research was performed based on a comparative study on fungal lipid production by a locally isolated strain Cunninghamella bainieri 2A1 in batch culture and repeated-batch culture using a nitrogen-limited medium. Lipid production in the batch culture was conducted to study the effect of different agitation rates on the simultaneous consumption of ammonium tartrate and glucose sources. Lipid production in the repeated-batch culture was studied by considering the effect of harvesting time and harvesting volume of the culture broth on the lipid accumulation. The batch cultivation was carried out in a 500 ml Erlenmeyer flask containing 200 ml of the fresh nitrogen-limited medium. Microbial culture was incubated at 30 °C under different agitation rates of 120, 180 and 250 rpm for 120 h. The repeated-batch culture was performed at three harvesting times of 12, 24 and 48 h using four harvesting cultures of 60%, 70%, 80% and 90%. Experimental results revealed that nitrogen source (ammonium tartrate) was fully utilized by C. bainieri 2A1 within 24 h in all agitation rates tested. It was also observed that a high amount of glucose in culture medium was consumed by C. bainieri 2A1 at 250 rpm agitation speed during the batch fermentation. Similar results showed that the highest lipid concentration of 2.96 g/L was obtained at an agitation rate of 250 rpm at 120 h cultivation time with the maximum lipid productivity of 7.0 × 10(-2) mg/ml/h. On the other hand, experimental results showed that the highest lipid concentration produced in the repeated-batch culture was 3.30 g/L at the first cycle of 48 h harvesting time using 70% harvesting volume, while 0.23 g/L gamma-linolenic acid (GLA) was produced at the last cycle of 48 h harvesting time using 80% harvesting volume.
Of the twenty microorganisms screened for metabolism of goniothalamin only Streptomyces aurofaciens ATCC 10762 and Nocardia species NRRL 5646 produced two metabolites, 3,4-dihydrogoniothalamin and 3,4,7,8 tetrahydrogoniothalamin. The identity of the isolated metabolites were established using TLC, HPLC, MS, IR, and 1H- and 13C-NMR spectroscopy. In addition, the substrate had been transformed into two unknown metabolites by Aspergillus niger ATCC 11394 and Septomyxa affinis ATCC 6737 in low yield. Three of the metabolites were also detected and identified in the urine and blood samples of the goniothalamin-treated Sprague-Dawley rats. The obtained results are in agreement with and support the principle of microbial models of mammalian metabolism.
Fungi is known to produce a wide range of biologically active metabolites and enzymes. Enzymes produced by fungi are utilized in food and pharmaceutical industries because of their rich enzymatic profile. Filamentous fungi are particularly interesting due to their high production of extracellular enzymes which has a large industrial potential. The aim of this study is to isolate potential soil fungi species that are able to produce functional enzymes for industries. Five Aspergillus species were successfully isolated from antibiotic overexposed soil (GPS coordinate of N3.093219 E101.40269) by standard microbiological method. The isolated fungi were identified via morphological observations and molecular tools; polymerase chain reactions, ITS 1 (5’- TCC GTA GGT GAA CCT GCG G3’) forward primer and ITS 4 (5’-TCC TCC GCT TAT TGA TAT GC-3’) reverse primer. The isolated fungi were identified as Aspergillus sydowii strain SCAU066, Aspergillus tamarii isolate TN-7, Aspergillus candidus strain KUFA 0062, Aspergillus versicolor isolate BAB-6580, and Aspergillus protuberus strain KAS 6024. Supernatant obtained via submerged fermentation of the isolated fungi in potato dextrose broth (PDB) and extracted via centrifugation was loaded onto specific media to screen for the production of xylanolytic, cellulolytic and amylolytic enzymes. The present findings indicate that Aspergillus sydowii strain SCAU066 and Aspergillus versicolor isolate BAB-6580 have great potential as an alternative source of xylanolytic, cellulolytic and amylolytic enzymes.
The influence of commercial prebiotics (fructo-oligosaccharides and inulin) and sugars (glucose and sucrose) on enhancing equol production from soymilk isoflavones by Bifidobacterium longum BB536 and Bifidobacterium breve ATCC 15700 was evaluated in vitro. Sterilized soymilk was inoculated with each bacterial species at 37 °C for 48 h. The growth and β-glucosidase enzyme activity for the two Bifidobacterium species in soymilk throughout fermentation were assessed. The highest viable count for B. breve (8.75 log CFU/ml) was reached at 36 h and for B. longum (8.55 log CFU/ml) at 24 h. Both bacterial species displayed β-glucosidase activity. B. breve showed increased enzyme activity (4.126 U) at 36 h, while B. longum exhibited maximum activity (3.935 U) at 24 h of fermentation. Among the prebiotics screened for their effect in isoflavones transformation to equol, inulin delivered the highest effect on equol production. The co-culture of B. longum BB536 and B. breve ATCC15700 in soymilk supplemented with inulin produced the highest level (11.49 mmol/l) of equol at 48 h of fermentation process. Level of daidzin declined whereas that of daidzein increased, and then gradually decreased due to formation of equol when soymilk was fermented using bifidobacterial. This suggests that the nutritional value of soymilk may be increased by increasing bioavailability of the bioactive ingredients. Collectively these data identify probiotics and prebiotic combinations suitable for inclusion in soymilk to enhance equol production.
In this study, Gelidium elegans is investigated for ethanol production. A combination of factors including different temperatures, acid concentration and incubation time was evaluated to determine the suitable saccharification conditions. The combination of 2.5% (w/v) H2SO4 at 120 °C for 40 min was selected for hydrolysis of the seaweed biomass, followed by purification, and fermentation to yield ethanol. The galactose and glucose were dominant reducing sugars in the G. elegans hydrolysate and under optimum condition of dilute acid hydrolysis, 39.42% of reducing sugars was produced and fermentation resulted in ethanol concentration of 13.27 ± 0.47 g/L. A modified method was evaluated for sample preparation for gas chromatography (GC) analysis of the ethanol content. A solvent mixture of acetonitrile and iso-butanol precipitated dissolved organic residues and reduced water content in GC samples at least by 90%. Results showed that this method could be successfully used for bioethanol production from seaweed.
Wild-cultivated medicinal mushroom Ganoderma lucidum was morphologically identified and sequenced using phylogenetic software. In submerged-liquid fermentation (SLF), biomass, exopolysaccharide (EPS) and intracellular polysaccharide (IPS) production of the identified G.lucidum was optimised based on initial pH, starting glucose concentration and agitation rate parameters using response surface methodology (RSM). Molecularly, the G. lucidum strain QRS 5120 generated 637 base pairs, which was commensurate with related Ganoderma species. In RSM, by applying central composite design (CCD), a polynomial model was fitted to the experimental data and was found to be significant in all parameters investigated. The strongest effect (p < 0.0001) was observed for initial pH for biomass, EPS and IPS production, while agitation showed a significant value (p < 0.005) for biomass. By applying the optimized conditions, the model was validated and generated 5.12 g/L of biomass (initial pH 4.01, 32.09 g/L of glucose and 102 rpm), 2.49 g/L EPS (initial pH 4, 24.25 g/L of glucose and 110 rpm) and 1.52 g/L of IPS (and initial pH 4, 40.43 g/L of glucose, 103 rpm) in 500 mL shake flask fermentation. The optimized parameters can be upscaled for efficient biomass, EPS and IPS production using G. lucidum.
The ability of oleaginous yeast Lipomyces starkeyi to efficiently produce lipids when cultivated on sap extracted from felled oil palm trunk (OPT) as a novel inexpensive renewable carbon source was evaluated. OPT sap was found to contain approximately 98 g/L glucose and 32 g/L fructose. Batch fermentations were performed using three different OPT sap medium conditions: regular sap, enriched sap, and enriched sap at pH 5.0. Under all sap medium conditions, the cell biomass and lipid production achieved were approximately 30 g/L and 60% (w/w), respectively. L. starkeyi tolerated acidified medium (initial pH ≈ 3) and produced considerable amounts of ethanol as well as xylitol as by-products. The fatty acid profile of L. starkeyi was remarkably similar to that of palm oil, one of the most common vegetable oil feedstock used in biodiesel production with oleic acid as the major fatty acid followed by palmitic, stearic and linoleic acids.
In recent years, many efforts have been directed to explore the methods to reduce the production costs of industrial lipase by improving the yield and the use of low-cost agricultural wastes. Coconut dregs, which is a lignocellulosic by-product from coconut oil and milk processing plants, is rich in cellulose (36%) and crude fat (9%). A newly isolated Bacillus stratosphericus has been demonstrated to perform cellulose hydrolysis on coconut dregs producing fermentable sugars. The highest extracellular lipase activity of 140 U/mL has been achieved in submerged fermentation with acid pre-treated coconut dregs. The lipase was found to be active over a wide range of temperatures and pHs. The activity of lipase can be generally increased by the presence of detergent ingredients such as Tween-80, cetyltrimethylammonium bromide, hydrogen peroxide and phosphate per sulphate. The great compatibility of lipase in commercial detergents has also underlined its potential as an additive ingredient in biodetergent formulations.
The removal of antibiotics and resistance genes in wastewater treatment plants has attracted widespread attention, but the potential role of residual antibiotics in the disposal of waste activated sludge (WAS) has not been clearly understood. In this study, the effect of roxithromycin (ROX) on volatile fatty acid (VFA) recovery from WAS anaerobic fermentation was investigated. The experimental results showed that ROX made a positive contribution to the production of VFAs. With the increase of ROX dosages from 0 to 100 mg/kg TSS, the maximum accumulation of VFAs increased from 295 to 610 mg COD/L. Mechanism studies revealed that ROX promoted the solubilization of WAS by facilitating the disruption of extracellular polymeric substances. In addition, ROX enhanced the activity of acetate kinase and inhibited the activities of α-glucosidase and coenzyme F420, and showed a stronger inhibitory effect on methane production than the hydrolysis process, thus resulting in an increase in VFA accumulation. These findings provide a new insight for the role of antibiotics in anaerobic fermentation of WAS.
This study was conducted to evaluate the potential of pineapple peel (PP) and pineapple crown leaves (PCL) as the substrate for vanillic acid and vanillin production. About 202 ± 18 mg L-1 and 120 ± 11 mg L-1 of ferulic acid was produced from the PP and PCL respectively. By applied response surface methodology, the ferulic acid yield was increased to 1055 ± 160 mg L-1 by treating 19.3% of PP for 76 min, and 328 ± 23 mg L-1 by treating 9.9% of PCL for 36 min in aqueous sodium hydroxide solution at 120 °C. The results revealed that PP extract was better than PCL extract for vanillic acid and vanillin production. Furthermore, the experiment also proved that large volume feeding was more efficient than small volume feeding for high vanillic acid and vanillin yield. Through large volume feeding, about 7 ± 2 mg L-1 of vanillic acid and 5 ± 1 mg L-1 of vanillin was successfully produced from PP extract via Aspergillus niger fermentation.
Simultaneous Saccharification and Fermentation (SSF) is a process where microbes have to first excrete extracellular enzymes to break polymeric substrates such as starch or cellulose into edible nutrients, followed by in situ conversion of those nutrients into more valuable metabolites via fermentation. As such, SSF is very attractive as a one-pot synthesis method of biological products. However, due to the co-existence of multiple biochemical steps, modeling SSF faces two major challenges. The first is to capture the successive chain-end and/or random scission of the polymeric substrates over time, which determines the rate of generation of various fermentable substrates. The second is to incorporate the response of microbes, including their preferential substrate utilization, to such a complex broth. Each of the above-mentioned challenges has manifested itself in many related areas, and has been competently but separately attacked with two diametrically different tools, i.e., the Population Balance Modeling (PBM) and the Cybernetic Modeling (CM), respectively. To date, they have yet to be applied in unison on SSF resulting in a general inadequacy or haphazard approaches to examine the dynamics and interactions of depolymerization and fermentation. To overcome this unsatisfactory state of affairs, here, the general linkage between PBM and CM is established to model SSF. A notable feature is the flexible linkage, which allows the individual PBM and CM models to be independently modified to the desired levels of detail. A more general treatment of the secretion of extracellular enzyme is also proposed in the CM model. Through a case study on the growth of a recombinant Saccharomyces cerevisiae capable of excreting a chain-end scission enzyme (glucoamylase) on starch, the interlinked model calibrated using data from the literature (Nakamura et al., Biotechnol. Bioeng. 53:21-25, 1997), captured features not attainable by existing approaches. In particular, the effect of various enzymatic actions on the temporal evolution of the polymer distribution and how the microbes respond to the diverse polymeric environment can be studied through this framework.
In this study, a selected γ-aminobutyric acid (GABA)-rich Malaysian strain Aspergillus oryzae NSK was collected from soy sauce koji. The strain was used to explore the effect of using renewable native sugar syrup, sugarcane, nipa, and molasses as fermentable substrates for developing a novel functional GABA soy sauce. We evaluated the strain using the chosen native sugars for 7 days using shake flask fermentation at 30 °C. The results showed optimum GABA concentration was achieved using cane molasses as the fermentable substrate (354.08 mg/L), followed by sugarcane syrup (320.7 mg/L) and nipa syrup (232.07 mg/L). Cane molasses was subsequently utilized as a substrate to determine the most suitable concentration for A. oryzae NSK to enhance GABA production and was determined as 50% g/L of glucose standard cane molasses. Our findings indicate that cane molasses can be used as a GABA-rich ingredient to develop a new starter culture for A. oryzae NSK soy sauce production.
In submerged-liquid fermentation, seven key parameters were assessed using one-factor-at-a-time to obtain the highest GABA yield using an industrial soy sauce koji Aspergillus oryzae strain NSK (AOSNSK). AOSNSK generated maximum GABA at 30 °C (194 mg/L) and initial pH 5 (231 mg/L), thus was able to utilize sucrose (327 mg/L of GABA) for carbon source. Sucrose at 100 g/L, improved GABA production at 646 mg/L. Single nitrogen sources failed to improve GABA production, however a combination of yeast extract (YE) and glutamic acid (GA) improved GABA at 646.78 mg/L. Carbon-to-nitrogen ratio (C8:N3) produced the highest cell (24.01 g/L) and GABA at a minimal time of 216 h. The key parameters of 30 °C, initial pH 5, 100 g/L of sucrose, combination YE and GA, and C8:N3 generated the highest GABA (3278.31 mg/L) in a koji fermentation. AOSNSK promisingly showed for the development of a new GABA-rich soy sauce.
Lipases are enzyme with versatile industrial applications can be produced by the solid-state fermentation (SSF) method and is an economical alternative for enzyme production assisted by fungus. In Malaysia, 5 million of copra waste were generated annually. Large amount of copra waste produced will cause an increasing amount of the waste dumped to the landfill. Copra waste is one of the potential substrates to produce lipase enzyme through SSF. Thus, the aim of this study is to optimize the lipase production by SSF associated by Aspergillus niger using the 23 full factorial design approach. In this study the factors affecting parameters that involved in the production of lipase enzyme such as temperature (25˚ and 35˚), substrates concentration (40% and 60%) and inoculum size of Aspergillus niger (1 and 9 petri dish) were determined. The maximum production of lipase was obtained after 120-hour incubation in SSF. The optimum condition for inoculum size of Aspergillus niger was 9 plates, 30°C of incubation temperature and 60 % moisture contents. The range of the concentration of lipase enzyme produced varied from 105 U/ml to 170 U/ml. When applied to the wastewater treatment, the reducing percentage of fat, oil and grease (FOG) in food processing wastewater is reduced from 219.4925mg/l to 169.467mg/l accounted to the amount of 34 % FOG removal. Lipase produced using copra waste as a substrate using SSF has the potential value to be developed in the future for various industry including wastewater treatment industry.
In this study, 4 Lactobacillus plantarum strains and 5 Lactobacillus fermentum strains adapting well to the unfavorable fruit system were isolated under different fruit environments. The fermentation ability of these autochthonous lactic acid bacteria (LAB) strains in blueberry juice, and the influence of microbial metabolism on juice composition were explored. After 48 h of fermentation, the viable cell counts exceeded 10.0 log CFU/mL, malic acid content decreased from 511.47 ± 10.50 mg/L to below 146.38 ± 3.79 mg/L, and lactic acid content increased from 0 mg/L to above 2184.90 ± 335.80 mg/L. Moreover, the metabolism of these strains exerted a profound influence on the phenolic composition of juice. Total phenolic content in blueberry juice increased by 6.1-81.2% under lactic acid fermentation, and the antioxidant capacity in vitro enhanced by at least 34.0%. Anthocyanin content showed a declining trend, while the profile of non-anthocyaninic phenolics exhibited complex changes. The increments of rutin, myricetin and gallic acid contents through 48 h lactic acid fermentation exceeded 136%, 71% and 38%, respectively. Instead, the contents of p-hydroxybenzoic acid and caffeic acid decreased with fermentation. Overall, Lactobacillus plantarum LSJ-TY-HYB-T9 and LSJ-TY-HYB-T7, and Lactobacillus fermentum LSJ-TY-HYB-C22 and LSJ-TY-HYB-L16 could be the suitable strains to produce fermented fruit juices, including blueberry in practical applications.
Yak dung is used as fuel in Tibetan homes; however, this use is hazardous to health. An alternative use of the dung that would be profitable and offset the loss as a fuel would be very beneficial. Sweet sorghum silage with yak dung biochar as an additive was compared with a control silage with no additives and three silages with different commercial additives, namely Lactobacillus buchneri, Lactobacillus plantarum and Acremonium cellulase. Biochar-treated silage had a significantly greater concentration of water-soluble carbohydrates than the other silages (76 vs 12.4-45.8 g/kg DM) and a greater crude protein content (75.5 vs 61.4 g/kg DM), lactic acid concentration (40.7 vs 27.7 g/kg DM) and gross energy yield (17.8 vs 17.4 MJ/kg) than the control silage. Biochar-treated and control silages did not differ in in vitro digestibility and in total gas (507 vs 511 L/kg DM) and methane production (57.9 vs 57.1 L/kg DM). Biochar inhibited degradation of protein and water-soluble carbohydrates and enhanced lactic acid production, which improved storability of feed. It was concluded that yak dung biochar is an efficient, cost-effective ensiling additive. The profit could offset the loss of dung as fuel and improve the health of Tibetan people.
Optimization of fermentation processes requires monitoring the species composition of starter cultures and their growth during fermentation. Most starter cultures contain closely related species. Nowadays, high-resolution melting (HRM) analysis is extensively used for multiplex identification of closely related species. In the present paper, we applied real-time polymerase chain reaction (PCR) with HRM analysis for the detection and differentiation of Lactobacillus sakei and L. curvatus. A primer pair was selected for the site of the rpoA gene of Lactobacillus spp. Eleven starter cultures and fifteen fermented sausages with a known bacterial composition were successfully tested using real-time PCR with HRM analysis with the developed primer pair.
Understanding the nature of ruminant nutrition and digestion is essential to improve feeding management and animal production. Among many approaches, manipulating ruminant nutrition and fermentation through feed supplementation is being practised and researched. Over the last decade, the utilization of vegetable oils in feed formulation and their effects on various aspects of ruminants have been reported by many researchers. It is important to understand the lipid metabolism in ruminants by microorganisms because it affects the quality of ruminant-derived products such as meat and milk. Majority of vegetable oil supplementation could reduce rumen protozoa population in ruminants due to the effects of medium-chain fatty acids (FAs). However, vegetable oil also contains unsaturated FAs that are known to have a negative effect on cellulolytic bacteria which could show inhibitory effects of the fibre digestion. In this paper, the physiology of nutrient digestion of ruminants is described. This paper also provides a current review of studies done on improvement and modification of rumen fermentation and microbial population through vegetable oil supplementation.
In this study polymerase chain reaction (PCR) was used to identify yeast in domestic ragi obtained
from two local markets in Sarawak and Pahang. These ragi are normally used as a dry starter in food fermentation (tapai) for Pahang (ST2) and Sarawak (ST3) and tuak (ST1) which is an alcoholic drink in Sarawak. Universal primer, NL1 and NL4 were used as a primer in this study to amplify D1/D2 fragment. Based on the result from the sequencing and after the BLAST search of the nucleotide sequences, the strain was confirmed as Candida glabrata (FN424108.) partial 26S rRNA gene, strain IMUFRJ 51955 for ST1, Saccharomyces cerevisiae(EU285514.1) isolate 35 26S ribosomal RNA gene, partial sequence for ST2 sample and Candida glabrata (FN393990.1) partial 26S rRNA gene, strain MUCL 51244 for ST3. All these strains were found in domestic ragi used for food fermentation.