The effect of pH and butyric acid supplementation on the production of butanol by a new local isolate of Clostridium acetobutylicum YM1 during batch culture fermentation was investigated. The results showed that pH had a significant effect on bacterial growth and butanol yield and productivity. The optimal initial pH that maximized butanol production was pH 6.0 ± 0.2. Controlled pH was found to be unsuitable for butanol production in strain YM1, while the uncontrolled pH condition with an initial pH of 6.0 ± 0.2 was suitable for bacterial growth, butanol yield and productivity. The maximum butanol concentration of 13.5 ± 1.42 g/L was obtained from cultures grown under the uncontrolled pH condition, resulting in a butanol yield (YP/ S ) and productivity of 0.27 g/g and 0.188 g/L h, respectively. Supplementation of the pH-controlled cultures with 4.0 g/L butyric acid did not improve butanol production; however, supplementation of the uncontrolled pH cultures resulted in high butanol concentrations, yield and productivity (16.50 ± 0.8 g/L, 0.345 g/g and 0.163 g/L h, respectively). pH influenced the activity of NADH-dependent butanol dehydrogenase, with the highest activity obtained under the uncontrolled pH condition. This study revealed that pH is a very important factor in butanol fermentation by C. acetobutylicum YM1.
MAIN CONCLUSION: X-ray microtomography results revealed that delignification process damaged the oil palm fibers, which correlated well with reduction of lignin components and increase of the phenolic content. Biodegradation investigation of natural fibers normally focuses on physico-chemical analysis, with less emphasis on physical aspect like fiber structures affect from microbial activity. In this work, the performance of Pycnoporus sanguineus to delignify oil palm empty fruit bunch fibers through solid-state fermentation utilizing various ratio of POME sludge was reported. In addition to tensile testing, physico-chemical and X-ray microtomography (µ-CT) analyses on the oil palm fibers were conducted to determine the effectiveness of the degradation process. The best ratio of fiber to fungi (60:40) was chosen based on the highest lignin loss and total phenolic content values and further investigation was performed to obtain fermentation kinetics data of both laccase and manganese peroxidase. µ-CT results revealed that delignification process damaged the pre-treated and untreated fibers structure, as evident from volume reduction after degradation process. This is correlated with reduction of lignin component and increase of the phenolic content, as well as lower stress-strain curves of the pre-treated fibers compared to the untreated ones (from tensile testing). It is suggested that P. sanguineus preferred to consume the outer layer of the fiber, before it penetrates through the cellular structure of the inner fiber.
Milk kefir fermentation has been used in households for generations. Consumption of milk kefir has been associated with various health benefits, presumably from the probiotics of yeast and bacteria that make up the kefir grains. In addition, many of the microbes are known to produce novel antimicrobial compounds that can be used for other applications. The microbes living inside kefir grains differ significantly depending on geographical location and production methods. In this study, we aimed to use metagenomic analysis of fermented milk by using three different kefir grains (kefir 1, kefir 2, and kefir 3) from different US sources. We analyzed the microbial compositions of the three milk fermentation samples. This study revealed that each sample contains unique and distinct groups of microbes, kefir 1 showed the least diversity, and kefir 3 showed the highest diversity. Kefir 3 is rich in Proteobacteria while kefir 2 is dominated by the Firmicutes. Using bacterial indicator growth analyses carried out by continuous readings from microplate-based bioreactor assays suggested that kefir 2 fermentation filtrate has higher antibacterial property. We have screened 30 purified cultures of kefir 2 sample and isolated two lactic acid bacteria strains with higher antibacterial activities; the two strains were identified as Leuconostoc mesenteroides 28-1 and Lentilactobacillus kefiri 25-2 by 16S genomic PCR with confirmed antibacterial activities of fermentation filtrate after growing under both aerobic and anaerobic conditions.
Pediococci are lactic acid bacteria (LAB) which have been used for centuries in the production of traditional fermented foods. There fermentative abilities were explored by the modern food processing industry in use of pediococci as starter cultures, enabling the production of fermented foods with distinct characteristics. Furthermore, some pediococci strains can produce bacteriocins and other antimicrobial metabolites (AMM), such as pediocins, which are increasingly being explored as bio-preservatives in various food matrices. Due to their versatility and inhibitory spectrum, pediococci bacteriocins and AMM are being extensively researched not only in the food industry, but also in veterinary and human medicine. Some of the pediococci were evaluated as potential probiotics with different beneficial areas of application associated with human and other animals' health. The main taxonomic characteristics of pediococci species are presented here, as well as and their potential roles and applications as starter cultures, as bio-preservatives and as probiotic candidates.
This study aims to investigate the impact of utilizing lactic acid fermentation (LAF) as storage method of food waste (FW) prior to dark fermentation (DF). LAF of FW was carried out in batches at six temperatures (4 °C, 10 °C, 23 °C, 35 °C, 45 °C, and 55 °C) for 15 days followed by biological hydrogen potential (BHP) tests. Different storage temperatures resulted in different metabolites distribution, with either lactate or ethanol being dominant (159.2 ± 20.6 mM and 234.4 ± 38.2 mM respectively), but no negative impact on BHP (averaging at 94.6 ± 25.1 mL/gVS). Maximum hydrogen production rate for stored FW improved by at least 57%. Microbial analysis showed dominance of lactic acid bacteria (LAB) namely Lactobacillus sp., Lactococcus sp., Weisella sp., Streptococcus sp. and Bacillus sp. after LAF. Clostridium sp. emerged after DF, co-existing with LAB. Coupling LAF as a storage method was demonstrated as a novel strategy of FW management for DF, for a wide range of temperatures.
The livestock industry needs to use crop straws that are highly digestible to improve feed productivity and reduce ruminal methane emissions. Hence, this study aimed to use the ozonation and pelleting processes to enhance the digestibility and reduce the ruminal methane emissions of wheat straw enriched with two nitrogen sources (i.e., urea and heat-processed broiler litter). Various analyses were conducted on the pellets, including digestibility indicators, mechanical properties, surface chemistry functionalization, chemical-spectral-structural features, and energy requirements. For comparison, loose forms of the samples were also analyzed. The nitrogen-enriched ozonated wheat straw pellets had 43.06 % lower lignin, 28.30 % higher gas production for 24 h, 12.28 % higher metabolizable energy, 13.78 % higher in vitro organic matter digestibility for 24 h, and 28.81 % higher short-chain fatty acid content than the nitrogen-enriched loose sample. The reduction of methane emissions by rumen microorganisms of nitrogen-enriched wheat straw by ozonation, pelleting, and ozonation-pelleting totaled 89.15 %, 23.35 %, and 66.98 %, respectively. The ozonation process resulted in a 64 % increase in the particle density, a 5.5-time increase in the tensile strength, and a 75 % increase in the crushing energy of nitrogen-enriched wheat straw. In addition, ozone treatment could also reduce the specific and thermal energy consumption required in the pelleting process by 15.10 % and 7.61 %, respectively.
In this study, strains producing feruloyl esterase were screened by Oxford Cup clear zones method and by evaluating the ability to decompose hydroxycinnamoyl esters. The strain was identified by 16S rDNA molecular biology. The contents of dietary fiber, reducing sugar, water-extractable arabinoxylans, phytic acid, total phenolics, total flavonoid, phenolic compounds composition, microstructure and antioxidant activity in bran before and after fermentation were studied. Eight strains producing feruloyl esterase were screened, among which strain P1 had the strongest ability to decompose hydroxycinnamoyl esters. The strain was identified and named L. fermentum NB02. Compared with unfermented bran, fermented bran exhibited higher contents of soluble dietary fiber, reducing sugar, water-extractable arabinoxylans, total phenolics, total flavonoid, and lower insoluble dietary fiber and phytic acid content. The dense surface structure of bran was destroyed, forming a porous structure. The release of phenolic compounds increased significantly. L. fermentum NB02 fermentation improved the antioxidant capacity of bran.
The effect of removal of resultant gas resulted in enhancement of the H2 yield. The technique of CO2 scavenging resulted in H2 yield being improved from 408 mL g(-1) to reach the maximum of 422 mL g'. The highest hydrogen productivity of 87.9 ml L(-1) h(-1) was obtained by CO2 scavenging. Biomass concentration was enhanced to 1.47 g L(-1), Y(P,X) of 287 ml g(-1) L(-1), Y(X/S) of 0.294 and Y(H2/s) of 0.0377 by the use of CO2 scavenging. The results suggested that the presence of the gaseous products in fermentation medium and headspace adversely effect biomass growth and hydrogen production.
Antioxidant properties of crude extract, partition extract, and fermented medium from Dendrobium sabin (DS) flower were investigated. The oven-dried DS flower was extracted using 100% methanol (w/v), 100% ethanol (w/v), and 100% water (w/v). The 100% methanolic crude extract showed the highest total phenolic content (40.33 ± mg GAE/g extract) and the best antioxidant properties as shown by DPPH, ABTS, and FRAP assays. A correlation relationship between antioxidant activity and total phenolic content showed that phenolic compounds were the dominant antioxidant components in this flower extract. The microbial fermentation on DS flower medium showed a potential in increasing the phenolic content and DPPH scavenging activity. The TPC of final fermented medium showed approximately 18% increment, while the DPPH of fermented medium increased significantly to approximately 80% at the end of the fermentation. Dendrobium sabin (DS) flower showed very good potential properties of antioxidant in crude extract and partition extract as well as better antioxidant activity in the flower fermented medium.
Biosurfactants are surface-active compounds produced by different microorganisms. The aim of this study was to introduce palm kernel cake (PKC) as a novel substrate for biosurfactant production using a potent bacterial strain under liquid state fermentation. This study was primarily based on the isolation and identification of biosurfactant-producing bacteria that could utilize palm kernel cake as a new major substrate. Potential bacterial strains were isolated from degraded PKC and screened for biosurfactant production with the help of the drop collapse assay and by analyzing the surface tension activity. From the screened isolates, a new strain, SM03, showed the best and most consistent results, and was therefore selected as the most potent biosurfactant-producing bacterial strain. The new strain was identified as Providencia alcalifaciens SM03 using the Gen III MicroPlate Biolog Microbial Identification System. The yield of the produced biosurfactant was 8.3 g/L.
The biosynthesis of biomedical products including lipid and gamma-linolenic acid (GLA) by Cunninghamella bainieri 2A1 was studied in repeated batch fermentation. Three key process variables, namely, glucose concentration, ammonium tartrate concentration, and harvesting time, were optimized using response surface methodology. Repeated batch fermentation was carried out by the cultivation of Cunninghamella bainieri 2A1 in nitrogen-limited medium with various nitrogen concentration (1-4 g/L) and glucose concentration (20-40 g/L) at three time intervals (12 h, 24 h, and 48 h). Experimental results showed that the highest lipid concentration of 6.2 g/L and the highest GLA concentration of 0.4 g/L were obtained in optimum conditions, where 20.2 g/L glucose, 2.12 g/L ammonium tartrate, and 48 h harvesting time were utilized. Statistical results showed that the interaction between glucose and ammonium tartrate concentration had highly significant effects on lipid and GLA biosynthesis (P < 0.01). Moreover, harvesting time had a significant interaction effect with glucose and ammonium tartrate concentration on lipid production (P < 0.05).
Oil palm frond (OPF) juice is a potential industrial fermentation substrate as it has high sugars content and the OPF are readily available daily. However, maximum sugars yield and storage stability of the OPF juice are yet to be determined. This study was conducted to determine the effect of physical pretreatment and storage duration of OPF petiole on sugars yield. Storage stability of OPF juice at different storing conditions was also investigated. It was found that OPF petiole squeezed by hydraulic pressing machine gave the highest sugars recovery at almost 40 g/kg, accounting for a recovery yield of 88%. Storage of OPF petiole up to 72 hrs prior to squeezing reduced the free sugars by 11 g/kg. Concentrated OPF juice with 95% water removal had the best storage stability at both 4 and 30°C, when it was stored for 10 days. Moreover, concentrated OPF syrup prepared by thermal processing did not give any Maillard effect on microbial growth. Based on our results, OPF juice meets all the criteria as a good fermentation substrate as it is renewable, consistently available, and easy to be obtained, it does not inhibit microbial growth and product formation, and it contains no impurities.
Ethanol was produced via the simultaneous saccharification and fermentation (SSF) of dilute sodium hydroxide treated corn stover. Saccharification was achieved by cultivating either Phanerochaete chrysosporium or Gloeophyllum trabeum on the treated stover, and fermentation was then performed by using either Saccharomyces cerevisiae or Escherichia coli K011. Ethanol production was highest on day 3 for the combination of G. trabeum and E. coli K011 at 6.68 g/100g stover, followed by the combination of P. chrysosporium and E. coli K011 at 5.00 g/100g stover. SSF with S. cerevisiae had lower ethanol yields, ranging between 2.88 g/100g stover at day 3 (P. chrysosporium treated stover) and 3.09 g/100g stover at day 4 (G. trabeum treated stover). The results indicated that mild alkaline pretreatment coupled with fungal saccharification offers a promising bioprocess for ethanol production from corn stover without the addition of commercial enzymes.
Volumetric mass transfer coefficient (kLa) is an important parameter in bioreactors handling viscous fermentations such as xanthan gum production, as it affects the reactor performance and productivity. Published literatures showed that adding an organic phase such as hydrocarbons or vegetable oil could increase the kLa. The present study opted for palm oil as the organic phase as it is plentiful in Malaysia. Experiments were carried out to study the effect of viscosity, gas holdup, and kLa on the xanthan solution with different palm oil fractions by varying the agitation rate and aeration rate in a 5 L bench-top bioreactor fitted with twin Rushton turbines. Results showed that 10% (v/v) of palm oil raised the kLa of xanthan solution by 1.5 to 3 folds with the highest kLa value of 84.44 h(-1). It was also found that palm oil increased the gas holdup and viscosity of the xanthan solution. The kLa values obtained as a function of power input, superficial gas velocity, and palm oil fraction were validated by two different empirical equations. Similarly, the gas holdup obtained as a function of power input and superficial gas velocity was validated by another empirical equation. All correlations were found to fit well with higher determination coefficients.
This research was carried out to evaluate the effects of flavone, myricetin, naringin, catechin, rutin, quercetin, and kaempferol at the concentration of 4.5% of the substrate (dry matter basis) on the rumen microbial activity in vitro. Mixture of guinea grass and concentrate (60 : 40) was used as the substrate. The results showed that all the flavonoids except naringin and quercetin significantly (P < 0.05) decreased the dry matter degradability. The gas production significantly (P < 0.05) decreased by flavone, myricetin, and kaempferol, whereas naringin, rutin, and quercetin significantly (P < 0.05) increased the gas production. The flavonoids suppressed methane production significantly (P < 0.05). The total VFA concentration significantly (P < 0.05) decreased in the presence of flavone, myricetin, and kaempferol. All flavonoids except naringin and quercetin significantly (P < 0.05) reduced the carboxymethyl cellulase, filter paperase, xylanase, and β -glucosidase activities, purine content, and the efficiency of microbial protein synthesis. Flavone, myricetin, catechin, rutin, and kaempferol significantly (P < 0.05) reduced the population of rumen microbes. Total populations of protozoa and methanogens were significantly (P < 0.05) suppressed by naringin and quercetin. The results of this research demonstrated that naringin and quercetin at the concentration of 4.5% of the substrate (dry matter basis) were potential metabolites to suppress methane production without any negative effects on rumen microbial fermentation.
The response surface method was applied in this study to improve cellulase production from oil palm empty fruit bunch (OPEFB) by Botryosphaeria rhodina. An experimental design based on a two-level factorial was employed to screen the significant environmental factors for cellulase production. The locally isolated fungus Botryosphaeria rhodina was cultivated on OPEFB under solid-state fermentation (SSF). From the analysis of variance (ANOVA), the initial moisture content, amount of substrate, and initial pH of nutrient supplied in the SSF system significantly influenced cellulase production. Then the optimization of the variables was done using the response surface method according to central composite design (CCD). Botryosphaeria rhodina exhibited its best performance with a high predicted value of FPase enzyme production (17.95 U/g) when the initial moisture content was at 24.32%, initial pH of nutrient was 5.96, and 3.98 g of substrate was present. The statistical optimization from actual experiment resulted in a significant increment of FPase production from 3.26 to 17.91 U/g (5.49-fold). High cellulase production at low moisture content is a very rare condition for fungi cultured in solid-state fermentation.
A comparative study on the stability and potential of alginate and pectin based beads for production of poultry probiotic cells using MRS medium in repeated batch fermentation was conducted. The bead cores, made of three types of materials, i.e., ca-alginate, ca-pectinate and ca-alginate/pectinate, were compared. The effect of single and double layer coatings using chitosan and core material, respectively, on the bead stability and cell production were also studied. The pectin based beads were found to be more stable than that of the alginate beads and their stability was further improved by coating with chitosan. The cell concentration in pectin based beads was comparable to that in the alginate beads. On the other hand, pectin based beads gave significantly lower cell concentration in the growth medium for the initial fermentation cycles when compared to the alginate beads. In conclusion, pectin was found to be potential encapsulation material for probiotic cell production owing to its stability and favourable microenvironment for cell growth.
Ethanolic fermentation using Saccharomyces cerevisiae was carried out on three types of hydrolysates produced from lignocelulosic biomass which are commonly found in Malaysia such as oil palm trunk, rubberwood and mixed hardwood. The effect of fermentation temperature and pH of hydrolysate was evaluated to optimize the fermentation efficiency which defined as maximum ethanol yield in minimum fermentation time. The fermentation process using different temperature of 25 degrees Celsius, 30 degrees Celsius and 40 degrees Celsius were performed on the prepared fermentation medium adjusted to pH 4, pH 6 and pH 7, respectively. Results showed that the fermentation time was significantly reduced with the increase of temperature but an adverse reduction in ethanol yield was observed using temperature of 40 degrees Celsius. As the pH of hydrolysate became more acidic, the ethanol yield increased. Optimum fermentation efficiency for ethanolic fermentation of lignocellulosic hydrolysates using S. cerevisiae can be obtained using 33.2 degrees Celsius and pH 5.3.
The kitchen waste fraction in municipal solid waste contains high organic matter particularly carbohydrate that can contribute to fermentable sugar production for subsequent conversion to bioethanol. This study was carried out to evaluate the influence of single and combination pretreatments of kitchen waste by liquid hot water, mild acid pretreatment of hydrochloric acid (HCl) and sulphuric acid (H2SO4) and enzymatic hydrolysis (glucoamylase). The maximum total fermentable sugar produced after combination pretreatment by 1.5% HCl and glucoamylase consisted of 93.25 g/L glucose, 0.542 g/L sucrose, 0.348 g/L maltose, and 0.321 g/L fructose. The glucose released by the combination pretreatment method was 0.79 g glucose/g KW equivalent to 79% of glucose conversion. The effects of the pre-treatment on kitchen waste indicated that the highest solubilization was 40% by the combination method of 1.5% HCl and glucoamylase. The best combination pre-treatment gave concentrations of lactic acid, acetic acid, and propionic acid of 11.74 g/L, 6.77 g/L, and 1.02 g/L, respectively. The decrease of aliphatic absorbance bands of polysaccharides at 2851 and 2923 cm(-1) and the increase on structures of carbonyl absorbance bands at 1600 cm(-1) reflects the progress of the kitchen waste hydrolysis to fermentable sugars. Overall, 1.5% HCl and glucoamylase treatment was the most profitable process as the minimum selling price of glucose was USD 0.101/g kitchen waste. Therefore, the combination pretreatment method was proposed to enhance the production of fermentable sugar, particularly glucose from kitchen waste as the feedstock for bioethanol production.