Inactivation of guanylate kinase in Bacillus sp. TL7-3 cultivated under an optimized ratio of carbon and nitrogen sources influenced GTP regeneration capability and sporulation.

Bacillus sp. TL7-3 has potential as a dietary supplement to promote human and animal health. It produces spores that can survive in harsh environments. Thus, when supplemented with nutrients, these spores can withstand the acidic pH of the stomach and resume vegetative development in the gut when exposed to growth-promoting conditions. Spores are formed as a cellular defense mechanism when a culture experiences stress and process optimization to achieve high spore production in a typical batch process remains challenging. Existing literature on the manipulation of gene expression and enzyme activity during batch cultivation is limited. Studies on the growth patterns, morphological changes, and relevant gene expression have aided in enhancing spore production. The present study used the response surface methodology for medium optimization. The model suggested that yeast extract and NH4Cl were significant factors controlling spore production. A comparison between the high weight ratio of carbon and nitrogen (C:N) substrates (8.57:1) in the optimized and basal media (0.52:1) showed an 8.76-fold increase in the final spore concentration. The expression of major genes, including codY, spo0A, kinA, and spo0F, involved in the sporulation was compared when cultivating Bacillus sp. TL7-3 in media with varying C:N ratios. At high C:N ratios, spo0A, kinA, and spo0F were upregulated, whereas codY was downregulated. This led to decreased guanylate kinase activity, resulting in a low guanosine triphosphate concentration and inactivation of CodY, thereby reducing the repression of spo0A and CodY-repressed genes and stimulating sporulation.

Microalgae growth-promoting bacteria for cultivation strategies: Recent updates and progress.

Microalgae growth-promoting bacteria (MGPB), both actinobacteria and non-actinobacteria, have received considerable attention recently because of their potential to develop microalgae-bacteria co-culture strategies for improved efficiency and sustainability of the water-energy-environment nexus. Owing to their diverse metabolic pathways and ability to adapt to diverse conditions, microalgal-MGPB co-cultures could be promising biological systems under uncertain environmental and nutrient conditions. This review proposes the recent updates and progress on MGPB for microalgae cultivation through co-culture strategies. Firstly, potential MGPB strains for microalgae cultivation are introduced. Following, microalgal-MGPB interaction mechanisms and applications of their co-cultures for biomass production and wastewater treatment are reviewed. Moreover, state-of-the-art studies on synthetic biology and metabolic network analysis, along with the challenges and prospects of opting these approaches for microalgal-MGPB co-cultures are presented. It is anticipated that these strategies may significantly improve the sustainability of microalgal-MGPB co-cultures for wastewater treatment, biomass valorization, and bioproducts synthesis in a circular bioeconomy paradigm.

The outlooks and key challenges in renewable biomass feedstock utilization for value-added platform chemical via bioprocesses.

The conversion of renewable biomass feedstock into value-added products via bioprocessing platforms has become attractive because of environmental and health concerns. Process performance and cost competitiveness are major factors in the bioprocess design to produce desirable products from biomass feedstock. Proper pretreatment allows delignification and hemicellulose removal from the liquid fraction, allowing cellulose to be readily hydrolyzed to monomeric sugars. Several industrial products are produced via sugar fermentation using either naturally isolated or genetically modified microbes. Microbial platforms play an important role in the synthesis of several products, including drop-in chemicals, as-in products, and novel compounds. The key elements in developing a fermentation platform are medium formulation, sterilization, and active cells for inoculation. Downstream bioproduct recovery may seem like a straightforward chemical process, but is more complex, wherein cost competitiveness versus recovery performance becomes a challenge. This review summarizes the prospects for utilizing renewable biomass for bioprocessing.

Successive process for efficient biovalorization of Brewers' spent grain to lignocellulolytic enzymes and lactic acid production through simultaneous saccharification and fermentation.

This study aimed to increase the value of brewers' spent grain (BSG) by using it as feedstock to produce lignocellulolytic enzymes and lactic acid (LA). Twenty-two fungal strains were screened for lignocellulolytic enzyme production from BSG. Among them, Trichoderma sp. showed the highest cellulase activity (35.84 ± 0.27 U/g-BSG) and considerably high activities of xylanase (599.61 ± 23.09 U/g-BSG) and ß-glucosidase (16.97 ± 0.77 U/g-BSG) under successive solid-state and submerged fermentation. The processes were successfully scaled up in a bioreactor. The enzyme cocktail was recovered and characterized. The maximum cellulase and xylanase activities were found at pH 5.0 and 50 °C, and the activities were highly stable at pH 4-8 and 30-50 °C. The enzyme cocktail was applied in simultaneous saccharification and fermentation of acid-pretreated BSG for LA production. The maximum LA obtained was 59.3 ± 1.0 g/L. This study has shown the efficient biovalorization of BSG, and this approach may also be applicable to other agro-industrial wastes.

Green biorefinery of shrimp shell waste for α-chitin and high-value co-products through successive fermentation by co-lactic acid bacteria and proteolytic fungus.

Green biorefinery process was conducted to extract α-chitin and high-value co-products from shrimp shell waste through microbial fermentation using mature coconut water (MCW) as a sole nutrient source. Symbiotic co-lactic acid fermentation (Co-LAF) by Lactobacillus plantarum and Streptococcus thermophilus produced higher levels of lactic acid (LA) and protease activity than their mono-cultures, which led to greater demineralization (DM) and deproteinization (DP) of shrimp shell powder (SSP). After optimizing Co-LAF through Response Surface Methodology and successive fermentation by an acid-active proteolytic fungus Rhizopus oligosporus, the highest DM of 94.0 ± 0.91 % and DP of 86.7 ± 0.1 % were achieved. Based on FT-IR, XRD, and SEM analysis, the bio-extracted chitin had similar structural characteristics to commercial α-chitin but with better quality. These strategies not only contribute to environmentally-friendly and cost-effective extraction of α-chitin (303 ± 18 mg/g-SSP), but also co-produce LA (57.18 ± 0.89 g/L), acid protease (4.33 ± 0.5 U/mL), bio-calcium (277 ± 12 mg-CaSO4/g-SSP), protein hydrolysate (268 ± 5 mg/g-SSP), and pigments (28.78 ± 1.56 µg/g-SSP).

Value-added green biorefinery co-products from ultrasonically assisted DES-pretreated Chlorella biomass.

This study pursued the goal of creating value-added co-products through an environmentally friendly biorefinery approach, employing ultrasonically assisted deep eutectic solvent (DES)-pretreated Chlorella biomass. The primary focus was on generating enriched biodiesel feedstock with exceptional fuel properties and developing hydroponic biofertilizer. The results demonstrated the effectiveness of a two-step process involving a 5-minute ultrasound-assisted DES pretreatment followed by ultrasound-assisted solvent extraction, which efficiently extracted lipids from Chlorella biomass, yielding biodiesel-quality lipids with good cetane number (59.42) and high heating value (40.11 MJ/kg). Notably, this two-step approach (78.04 mg-lipid/g-microalgal biomass) led to a significant 2.10-fold increase in lipid extraction compared to a one-step process (37.15 mg-lipid/g-microalgal biomass) that combined ultrasound-assisted DES pretreatment and solvent extraction. Importantly, the aqueous extract derived from lipid-extracted microalgal biomass residues (LMBRs) showed promise as a component in hydroponic biofertilizer production, supporting lettuce growth in hydroponic deep water culture system. Consequently, microalgae biorefinery co-products hold tremendous potential in enhancing the profitability and sustainability of interconnected sectors, encompassing renewable energy, agriculture, and the environment.

Microalgae as tools for bio-circular-green economy: Zero-waste approaches for sustainable production and biorefineries of microalgal biomass.

Microalgae are promising organisms that are rapidly gaining much attention due to their numerous advantages and applications, especially in biorefineries for various bioenergy and biochemicals. This review focuses on the microalgae contributions to Bio-Circular-Green (BCG) economy, in which zero-waste approaches for sustainable production and biorefineries of microalgal biomass are introduced and their possible integration is discussed. Firstly, overviews of wastewater upcycling and greenhouse gas capture by microalgae are given. Then, a variety of valuable products from microalgal biomass, e.g., pigments, vitamins, proteins/peptides, carbohydrates, lipids, polyunsaturated fatty acids, and exopolysaccharides, are summarized to emphasize their biorefinery potential. Techno-economic and environmental analyses have been used to evaluate sustainability of microalgal biomass production systems. Finally, key issues, future perspectives, and challenges for zero-waste microalgal biorefineries, e.g., cost-effective techniques and innovative integrations with other viable processes, are discussed. These strategies not only make microalgae-based industries commercially feasible and sustainable but also reduce environmental impacts.

Promoting Magnusiomyces spicifer AW2 Cell-Bound Lipase Production by Co-culturing with Staphylococcus hominis AUP19 and Its Application in Solvent-Free Biodiesel Synthesis.

Yeast-bacterium interaction has recently been investigated to benefit the production of cell-bound lipases (CBLs). Staphylococcus hominis AUP19 supported the growth of Magnusiomyces spicifer AW2 in a palm oil mill effluent (POME) medium to produce CBLs through a bioremediation approach, including oil and grease (O&G) and chemical oxygen demand (COD) removals. This research used the yeast-bacterium co-culture to optimize CBLs and cell biomass (CBM) productions through bioremediation using the statistical Plackett-Burman design and response surface methodology-central composite design. The CBLs were finally applied in biodiesel synthesis. The CBM of 13.8 g/L with CBLs activity at 3391 U/L was achieved after incubation at room temperature (RT, 30 ± 2 °C) for 140 h in 50% POME medium, pH 7.0, containing 1.23% (w/v) ammonium sulfate. Bacterium promoted yeast growth to achieve bioremediation with 87.9% O&G removal and 84.5% COD removal. Time course study showed that the CBLs activity was highest at 24 h cultivation (4103 U/L) and retained 80% and 60% of activities at 4 °C and RT after 5 weeks of storage. The CBLs application successfully yielded 77.3% biodiesel from oleic acid (esterification) and 86.4% biodiesel from palm oil (transesterification) within 72 h in solvent-free systems. This study highlights that yeast-bacterium co-culture and POME should receive more attention for potential low-cost CBLs production through bioremediation, i.e., O&G and COD removals, while the CBLs as biocatalysts are promising for significant contribution to an effective strategy for economic green biodiesel production.

Potential of butanol production from Thailand marine macroalgae using Clostridium beijerinckii ATCC 10132-based ABE fermentation.

The economical bio-butanol-based fermentation process is mainly limited by the high price of first-generation biomass, which is an intensive cost for the pretreatment of second-generation biomass. As third-generation biomass, marine macroalgae could be potentially advantageous for conversion to clean and renewable bio-butanol through acetone-butanol-ethanol (ABE) fermentation. In this study, butanol production from three macroalgae species (Gracilaria tenuistipitata, Ulva intestinalis, and Rhizoclonium sp.) by Clostridium beijerinckii ATCC 10132 was assessed comparatively. The enriched C beijerinckii ATCC 10132 inoculum produced a high butanol concentration of 14.07 g L-1 using 60 g L-1 of glucose. Among the three marine seaweed species, G. tenuistipitata exhibited the highest potential for butanol production (1.38 g L-1 ). Under the 16 conditions designed using the Taguchi method for low-temperature hydrothermal pretreatment (HTP) of G. tenuistipitata, the maximum reducing sugar yield rate of 57.6% and ABE yield of 19.87% were achieved at a solid to liquid (S/L) ratio of 120, temperature of 110°C, and holding time of 10 min (Severity factor, R0 1.29). In addition, pretreated G. tenuistipitata could be converted to 3.1 g L-1 of butanol using low-HTP at an S/L ratio of 50 g L-1 , temperature of 80°C (R0 0.11), and holding time of 5 min.

Enhanced biovalorization of palm biomass wastes as biodiesel feedstocks through integrated solid-state and submerged fermentations by fungal co-cultures.

Palm empty fruit bunches (EFB) were valorized into fungal lipids by oleaginous fungus Aspergillus tubingensis TSIP9 under solid-state fermentation (SSF) and submerged fermentation (SmF). An integrated SSF-SmF process increased lipid production from 116.2 ± 0.1 mg/g-EFB under SSF and 60.1 ± 0.2 under SmF up to 124.9 ± 0.5 mg/g-EFB, possibly due to the combined benefits of dispersed mycelia forming during SSF and better mass transfer during SmF. As A. tubingensis lacks sufficient ß-glucosidase, it was co-cultured with high ß-glucosidase-producing Trichoderma reesei QM 9414. The co-cultures improved overall lipid yields likely due to synergistic interaction of the two fungi. After inoculum size was optimized and the co-cultures were performed in bioreactors, the lipid yield was increased up to 205.1 ± 1.1 mg/g-EFB. The fatty acid composition of fungal lipids indicated their potential use as biodiesel feedstocks. The fungal fermentation of EFB also provided cellulose pulp residues. These strategies could be practical options for low-cost biovalorization of biomass wastes.

Chitosan-coated oleaginous microalgae-fungal pellets for improved bioremediation of non-sterile secondary effluent and application in carbon dioxide sequestration in bubble column photobioreactors.

Oleaginous microalga Scenedesmus sp. SPP was rapidly immobilized in oleaginous fungal pellets by their opposite-surface-charges. Microalgae-fungal (MF) pellets were more effective in bioremediation of non-sterile secondary effluent than mono-culture. The optimal hydraulic retention time for dual bioremediation in semi-continuous mode was 72 h. The MF pellets coated with 0.4 %-chitosan improved removal efficiencies of COD, total nitrogen (TN), and total phosphorus (TP) up to 96.2±0.0 %, 88.2±2.8 % and 71.5±0.7 %, respectively, likely because of better cell retention and more nutrient adsorption and assimilation. Dual bioremediation by coated MF pellets was also successfully scaled up in 30-L bubble-column photobioreactors with improved COD, TN, and TP removal efficiencies of 98.5±0.0 %, 90.2±0.0 % and 79.5±2.1 %, respectively. This system also effectively removed CO2 from simulated flue gas at 71.2±0.4 % and produced biomass with high lipid content. These results highlight the effectiveness of bio-immobilization by fungal pellets; chitosan coating; and their practical applications in bioremediation and CO2 sequestration.

Isolation and screening of microorganisms for high yield of succinic acid production.

This study involves the isolation of succinic acid (SA)-producing microorganisms from different samples, including the rumen, sludge, soil, and wastewater. For primary screening, 29 isolates exhibited a zone of clearance around the colony, indicating acid production. For secondary screening using thin-layer chromatography, only two isolates symbolized SA production according to their Rf values. These two isolates were further identified as Bacillus velezensis and Enterococcus gallinarum by phylogenetic analysis using the neighbor-joining method. The high SA concentrations of 50.2 and 66.9 g/L were produced by B. velezensis and E. gallinarum with an SA yield of 0.836 and 1.12 g/g glucose, respectively. The high SA concentration from these newly isolated strains was achieved with a low formation of unwanted acids compared with those from Actinobacillus succinogenes ATCC 55618. Moreover, E. gallinarum was cultured in palm oil mill wastewater (POMW) and molasses, which were cheap substrates. The high SA production of 73.9 g/L with low other acids (the ratio of SA to total acids = 0.917) was achieved using POMW and molasses (80:20) as substrates.

Solid-state fermentation of Saba banana peel for pigment production by Monascus purpureus.

Eco-friendly natural pigment demand has ever-increasing popularity due to health and environmental concerns. In this context, the aim of this study was to evaluate the feasibility use of Saba banana peel as low-cost fermentable substrate for the production of pigments, xylanase and cellulase enzymes by Monascus purpureus. Among the strains tested, M. purpureus TISTR 3385 produced pigments better and had higher enzyme activities. Under the optimal pigment-producing conditions at the initial moisture content of 40% and initial pH of 6.0, the pigments comprising yellow, orange, and red produced by the fungi were achieved in the range of 0.40-0.93 UA/g/day. The maximum xylanase and cellulase activities of 8.92 ± 0.46 U/g and 4.72 ± 0.04 U/g were also obtained, respectively. More importantly, solid-state fermentation of non-sterile peel could be achieved without sacrificing the production of the pigments and both enzymes. These indicated the potential use of the peel as fermentable feedstock for pigment production by the fungi and an environmental-friendly approach for sustainable waste management and industrial pigment and enzyme application.

Deploying two-stage anaerobic process to co-digest greasy sludge and waste activated sludge for effective waste treatment and biogas recovery.

High-strength waste activated sludge (WAS) and greasy sludge (GS) were largely generated from canned tuna processing. This study reports the performance of the two-stage anaerobic process for co-digesting WAS and GS. Various WAS:GS mixing ratios of 0:100, 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, and 100:00 (volatile solids (VS) basis) were investigated in batch acidogenic stage at ambient (30 °C ± 3 °C), 55 °C, and 60 °C temperatures. Subsequently, the effluents from the first stage were used to produce methane in the second methanogenic stage at an ambient temperature. The highest methane yield of 609 mL CH4/g-VSadded was achieved using acidogenic effluents generated from a WAS:GS mixing ratio of 40:60 at an ambient temperature. The first-order kinetic constants (k) for the first (k1) and second (k2) stages were subsequently estimated to be 0.457 d-1 and 0.139 d-1, respectively. The obtained k constants were further used to predict the hydraulic retention time (HRT) for the two continuously stirred tank reactors (CSTR) in series. Consequently, the calculated 4-day HRT and 20-day HRT for 50-L CSTR1 and 250-L CSTR2, respectively, were used to operate the continuous two-stage process at an ambient temperature by feeding with a 40:60-WAS:GS mixing ratio. A satisfactory methane yield of 470-mL CH4/g-VS along with 75% chemical oxygen demand (COD) removal was generated. Furthermore, the predicted methane yield of 450-mL CH4/g-VS obtained from the simple kinetic CSTR model resembled the experimental yield with 96% accuracy. The obtained experimental results demonstrate that WAS and GS co-digestion could be successfully accomplished using a practical two-stage anaerobic process operated at an ambient temperature.

Dual-bioaugmentation strategy to enhance the formation of algal-bacteria symbiosis biofloc in aquaculture wastewater supplemented with agricultural wastes as an alternative nutrient sources and biomass support materials.

This study performs an integrated evaluation of the formation and distribution of algal-bacterial bioflocs in aquaculture wastewater supplemented with agricultural waste, together with an assessment of their behavior in the microbial community and of the water quality of the system in which a new bioaugmentation strategy was applied. Results indicated that the dual bioaugmentation strategy via the consortium addition of bacteria and microalgae had the highest formation performance, providing the most compact biofloc structure (0.59 g/L), excellent settleability (71.91%), and a large particle diameter (4.25 mm). The fed-batch supplementation of molasses and rice bran, in terms of changes in the values of COD, NH4+, NO3-, and PO43-, stimulated the formation of biofloc through algal-bacterial bioflocs and microbe-rice bran complexes within a well-established microbial community. These findings provide new insight into the influence of bioaugmentation on the formation of an innovative algal-bacterial biofloc.

Performance of pilot scale two-stage anaerobic co-digestion of waste activated sludge and greasy sludge under uncontrolled mesophilic temperature.

Waste-activated sludge (WAS) and greasy sludge (GS) discharged from the canned tuna industry are considerably characterized as harsh organic wastes to be individually treated by using traditional anaerobic digestion. This study was attempted to anaerobically co-digest WAS and GS in continuous pilot scale two-stage process, comprising the first 50 L continuous stir tank reactor (CSTR1) and the second 250 L continuous stir tank reactor (CSTR2). The two-stage co-digesting operation of dewatered WAS:GS ratio of 0.4:1 (g-VS) at ambient temperature with the organic loading rate (OLR) of 12.6 ± 0.75 g-VS/L·d and 2.26 ± 0.13 g-VS/L·d, corresponding to 3-day and 17-day hydraulic retention time (HRT) for the first and second stage, respectively generated highest methane production rate of 957 ± 86 mL-CH4/L·d, corresponding to methane yield of 423.4 ± 36 mL-CH4/g-VS. Organic removal efficiency obtained was around 67.5% on COD basis. The microbial diversity was depended on the process's activity. Bacteria were mostly detected in the CSTR1, dominating with the phylum Firmicutes and Proteobacteria, whereas genus Methanosaeta archaea were found dominantly in the CSTR2. The economic analysis of process shows payback period (PBP), internal rate of return (IRR), and net present value (NPV) of 3 years, 30%, and 250,177 USD, respectively. This study demonstrated the potential approach to applying the two-stage anaerobic co-digestion process to stabilize both WAS and GS along with generating valuable bioenergy carriers.

Lipid Profile, Antioxidant and Antihypertensive Activity, and Computational Molecular Docking of Diatom Fatty Acids as ACE Inhibitors.

Diatoms, as single cell eukaryotic microalgae, are rich sources of lipids, which have either beneficial or detrimental effects on the prevention and treatment of many diseases. Gas chromatography-mass spectrometry (GC-MS) identified diatom lipids with high levels of essential fatty acids (EFAs), especially polyunsaturated FAs (PUFAs) containing both omega-3 and omega-6. Nutritional values of FAs indicated possible applications in the pharmaceutical, nutraceutical, and functional food industries. Diatom FAs showed antioxidative potential on harmful radicals by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) scavenging, with high inhibition of the angiotensin-converting enzyme (ACE) that causes cardiovascular disease (CVD) and hypertension. A computational molecular docking simulation confirmed the inhibition mechanisms of FAs on ACE, with comparable levels of binding free energy to chemically synthesized ACE drugs. Findings suggested that diatom lipids showed potential for use as alternative ACE inhibitors or food supplement for CVD prevention.

Application of palm oil mill waste to enhance biogas upgrading and hornwort cultivation.

The potential of oil palm ash (OPA) to enhance H2S and CO2 removal from biogas by scrubbing with maturation pond effluent (MPE), and further the treatment of biogas scrubber effluent (BSE) by Ceratophyllum demersum L. (hornwort) cultivation were investigated in this study. The results show that OPA + MPE solution with pH 9.3 and alkalinity 7525 mg CaCO3/L was obtained with 0.7 kg/L OPA loading. A pilot scale scrubber was used to study the effects of absorbent flow rates of 60-210 L/h on upgrading to 300 L/h field biogas stream. At 210 L/h, the CO2 removal efficiencies were 33% and 53% for MPE and OPA + MPE, respectively. To approach 100% H2S removal efficiency, the minimum flow rates were 120 L/h for MPE and 90 L/h for OPA + MPE. 50-150 g wet weight of hornwort in 30 L diluted POME were loaded to investigate appropriate initial hornwort loading level for hornwort cultivation. The highest specific growth rate of 0.045 day-1 with biomass production of 3.8 g/day were obtained with a 50 g initial loading. Among the wastewaters (MPE, OPA + MPE, and BSE) treatment using hornwort cultivation, the highest 0.035 day-1 specific growth rate and 2.6 g/day biomass production of hornwort were obtained in diluted BSE cultivation, and in 3 weeks of cultivation. COD, nitrate, phosphate, and alkalinity decreased by 76%, 76%, 55%, and 5%, respectively. The Eco-Efficiency concept for palm oil mill waste utilization proposed in this study has a high potential for enhanced biogas upgrading by using OPA + MPE, and hornwort is a good candidate for BSE post-treatment integrated with biomass production.

Potential use of industrial by-products as promising feedstock for microbial lipid and lipase production and direct transesterification of wet yeast into biodiesel by lipase and acid catalysts.

This work attempted the conversion of crude glycerol to lipid and lipase by Yarrowia lipolytica and the direct transesterification of wet yeast by its lipase into biodiesel via response surface methodology to enhance the cost-effectiveness of biodiesel production from the lipids. The yeast grew better and accumulated a high amount of lipids on the waste combined with fish waste hydrolysate, but only exhibited high lipase activity on the waste supplemented with surfactants (i.e., gum Arabic, Tween 20, Tween 80). However, the combination of both wastes and Tween 80 further improved growth, lipid productivity, and lipase activity. More importantly, lipase-direct transesterification under optimal conditions (wet cell concentration of 17.97 mg-DCW, methanol loading of 8.21 µL, and hexane loading of 10.26 µL) followed by acid-catalyst transesterification (0.4 M H2SO4), offered high FAME yields (>90%), showing the efficiency of the process when applied for the industrialization of biodiesel production from microbial lipids.

Impact of environmental factors on Streptomyces spp. metabolites against Botrytis cinerea.

Botrytis cinerea is an economically important disease on numerous vegetables including tomato. From our previous studies, a spore suspension of Streptomyces philanthi RL-1-178 and RM-1-138 and Streptomyces mycarofaciens SS-2-243 showed strong inhibition against B. cinerea. In this study, the efficacy of their antifungal metabolites against B. cinerea was investigated after enhancing the production through the optimum culture medium and environmental conditions (temperature, light/dark cycle). In vitro studies indicated that glucose yeast-malt (GYM) agar and incubation at 28°C were optimal for growth and mass spore production of all three Streptomyces strains. Moreover, light/dark conditions had a positive effect on the growth and spore production of S. philanthi RM-1-138 and RL-1-178 but not on S. mycarofaciens SS-2-243. Both strains of S. philanthi possessed an antifungal activity against B. cinerea (100% inhibition) while S. mycarofaciens showed different results on PDA (83% inhibition) and GYM (88% inhibition) at the optimum incubation temperature at 21°C. The antifungal compounds from S. philanthi RM-1-138 exhibited the highest protection efficacy against B. cinerea on tomato leaves (82.89% and 0.33 cm2 lesion areas symptoms). The antifungal compounds RM-1-138, identified by GC-MS, were greatly altered based on components concentration under various temperatures and light/dark conditions. The anti-B. cinerea of S. philanthi RM-1-138 was established at a higher level in several metabolic compounds in the dark condition (11 and 32 antifungal compounds after incubation at 21°C and 28°C, respectively) than in the light condition (11 and 19 antifungal compounds after incubation at 21°C and 28°C, respectively). At 21°C, the dominant component was acetic acid (67.41% and 68.77% in light and dark conditions, respectively) while at 28°C, benzeneacetamide (43.58% in light) and propanamide (20.68% in the dark) were dominant. The results clearly demonstrated the significant influence of environmental factors on the production of antifungal metabolites of Streptomyces spp.