Coproduction of inulinase and invertase by Galactomyces geotrichum in whey-based medium and evaluation of additional nutrients.

The purpose of this research was to evaluate the suitability of whey as an effective medium for the coproduction of inulinase and invertase by an oleaginous yeast Galactomyces geotrichum and to investigate the effects of some additional carbon and nitrogen sources. The nutritional factors and composition of the medium have a great impact on the production pathways of microbial enzymes. To deepen the research, a Taguchi design was employed to quickly scan the best conditions. First, the cheese whey was partly deproteinized and investigated as the sole medium for the yeast. The next step was performed to study the effects of inulin, sucrose and lactose as carbon sources and ammonium sulfate, yeast extract and casein as nitrogen sources. All analyses (Taguchi and ANOVA) were performed using Minitab software. Whey-based medium without any additional carbon and nitrogen sources gave inulinase and invertase activities as 54.6 U/mL and 47.4 U/mL, respectively. Maximum inulinase activity was obtained as 77.9 U/mL using inulin as the carbon source without any nitrogen source. The highest I/S ratio was found as 2.08. On the other hand, the highest invertase activity was carried out as 50.85 U/mL in whey-based medium using lactose as carbon source without any additional nitrogen source. This is the first report about partly deproteinized whey-based medium utilization for simultaneous inulinase and invertase production by G. geotrichum TS-61. Moreover, the effects of carbon and nitrogen sources were investigated in detail.

Whey is a sufficient medium for inulinase and invertase productionInulin is an excellent carbon source for enhanced inulinase activityTaguchi orthogonal array presents an effective and quick screening method for the fermentation process.

Starch-based antifungal edible coatings to control sour rot caused by Geotrichum citri-aurantii and maintain postharvest quality of 'Fino' lemon.

BACKGROUND: Two edible coating (EC) emulsions based on potato starch (F6 and F10) alone or formulated with sodium benzoate (SB, 2% w/w) (F6/SB and F10/SB) were evaluated to maintain postharvest quality of cold-stored 'Fino' lemons and control sour rot on lemons artificially inoculated with Geotrichum citri-aurantii. Previous research showed the potential of these ECs to improve the storability of 'Orri' mandarins and reduce citrus green and blue molds caused by Penicillum digitatum and Penicillium italicum, respectively. RESULTS: The coatings F6/SB and F10/SB significantly reduced sour rot incidence and severity compared to uncoated control samples on lemons incubated at 28 °C for 4 and 7 days. The F6/SB coating reduced weight loss and gas exchange compared to uncoated fruit after 2 and 4 weeks of storage at 12 °C plus a shelf life of 1 week at 20 °C, without adversely affecting the lemon physicochemical quality. CONCLUSION: Overall, the F6/SB coating formulation, composed of pregelatinized potato starch, glyceryl monostearate, glycerol, emulsifiers and SB, with a total solid content of 5.5%, showed the best results in reducing citrus sour rot and maintaining the postharvest quality of cold-stored 'Fino' lemons. Therefore, it showed potential as a new cost-effective postharvest treatment suitable to be included in integrated disease management programs for citrus international markets with zero tolerance to chemical residues. © 2021 Society of Chemical Industry.

Isolation, identification and optimization of enhanced production of laccase from Galactomyces geotrichum under solid-state fermentation.

Laccases are a group of oxidases that catalyze the oxidation of a wide range of electron rich substrates like phenolic compounds, lignin and aromatic amines. They are of interest because of their potential to be used in environmental and industrial applications. In this research, potent laccase producer fungi were screened and isolated from olive mill wastewater (OMW). One of the 23 isolated fungi was identified as Galactomyces geotrichum based on 18S rDNA sequence analysis that detected good laccase activity. Produced laccase had a molecular weight of 55 kDa that was confirmed by zymogram analysis. This is the first report about the optimization of laccase Production by G. geotrichum under solid-state fermentation. The optimization was made by the Taguchi design of experiments (DOE) methodology. An orthogonal array (L25) was designed using Minitab 19 software to study four effective process factors in five levels for laccase production. The optimum condition derived was; moisture content (80%), fermentation time (14 day), CuSO4⋅5H2O as the inducer (300 µM), glucose as a co-substrate (5 g/L). Maximum laccase activity of 52.86 (U/g of dry substrate) was obtained using optimum fermentation condition. This study aimed to better understand the laccase producing microorganisms in OMW and take them to OMW treatment that is rich in phenolic compounds.

Curative activity of postharvest GRAS salt treatments to control citrus sour rot caused by Geotrichum citri-aurantii.

The effectiveness of the "generally recognized as safe" (GRAS) salts potassium sorbate (PS), sodium benzoate (SB), sodium ethylparaben (SEP) and sodium methylparaben (SMP) to control sour rot, caused by Geotrichum citri-aurantii, was assessed by dipping economically important citrus species and cultivars in aqueous solutions for 30, 60 or 150 s at 20 °C, followed by examination after 8 d of storage at 28 °C. Curative activity was determined because the fruit were inoculated 24 h prior to treatment. Dipping fruit for 60 s in SMP (200 mM), SEP (200 mM) or SB (3% w/v) were very effective and reduced sour rot incidence and severity by up to 90%. Their effectiveness was similar or superior to that of the conventional fungicide propiconazole (PCZ). In contrast, PS (200 mM) did not control sour rot on 'Oronules' or 'Ortanique' mandarins, but it reduced sour rot incidence on 'Barnfield' oranges by 50% compared to inoculated, water-treated control fruit. Sour rot was better controlled on oranges than on mandarins. Furthermore, heating the solutions to 50 °C enhanced their effectiveness, while post-treatment rinsing of the fruit with tap water reduced their effectiveness. Dipping 'Valencia Late' oranges in SB (3% w/v) or SMP (200 mM) for 60 s followed by long storage for up to 8 weeks at 5 °C and 90% RH, reduced sour rot incidence from 55% among water-treated control fruit to 2 to 6%, and matched the effectiveness of PCZ. No fruit in any test were visibly harmed. Both SB and SMP salts could be potential alternatives to conventional fungicides, such as PCZ or guazatine, for the integrated postharvest management of citrus sour rot.

Production of volatile compounds by yeasts using hydrolysed grape seed oil obtained by immobilized lipases in continuous packed-bed reactors.

Lipases CAL-B, TLL, and RML were used in the synthesis of free fatty acids of grape seed oil as heterogeneous substrate. The best enzyme was used to optimize the reaction variables temperature, enzyme content, and molar ratio of water:oil in batch reactions using experimental planning. The ideal conditions to produce free fatty acids using pure RML were 45 °C, 12:1 substrate molar ratio, and 15% enzyme, resulting in 66% of oil hydrolysis and a productivity of 0.54 mol L-1 min-1 in 4 h of reaction at 180 rpm. Repeated batches of reaction were performed testing the operational stability of RML, results showing that this enzyme could be used for at least 20 cycles keeping more than 80% of its initial activity, suggesting its potential use in industrial processes. The synthesis of free fatty acids was then evaluated in continuous reactions using packed-bed reactor (PBR). The highest productivity in the continuous process was 6.85 mol L-1 min-1, using only RML, showing an operational stability higher than 80% of its initial conversion capacity after 11 days of operation, at a flow rate of 0.13 mL min-1 at 45 °C. We evaluated the use of this hydrolyzed oil as substrate for lactone bioproduction using Galactomyces geotrichum UFMG-CM-Y3276, G. geotrichum UFMG-CM-Y3558, and Geotrichum klebahnii UFMG-CM-Y3014 screened for their oil-hydrolysis ability. Volatile compounds were qualitatively identified in GC-MS as γ-octalactone and γ-nonalactone.

Biocontrol ability and action mechanism of Metschnikowia citriensis against Geotrichum citri-aurantii causing sour rot of postharvest citrus fruit.

This study investigated the biocontrol efficiency of Metschnikowia citriensis strain FL01 against Geotrichum citri-aurantii, and evaluated possible mechanisms. The results showed that M. citriensis could effectively control the development of sour rot, and significantly inhibit the mycelial growth and spore germination of G. citri-aurantii. The population dynamics results and Scanning electron microscopy (SEM) analysis indicated that M. citriensis could rapidly colonize wounds and tightly adhere to the surface of the wounds to compete with G. citri-aurantii for nutrition and space. M. citriensis also showed the biofilm formation action in vitro. The response of G. citri-aurantii to different components of M. citriensis culture showed that only the yeast cells but not the extracellular metabolites and the volatile organic compounds (VOCs) exhibited inhibitory effect on the growth of G. citri-aurantii. M. citriensis adhered to the hyphae of G. citri-aurantii loosely and sparsely, and the production of lytic enzymes ß-1, 3-glucanase (GLU) and Chitinase (CHI) could not be induced by G. citri-auranti. Iron affected the pulcherrimin pigment production and antagonism of M. citriensis indicating iron depletion as the most important antagonistic mechanism. Besides, M. citriensis also induced resistance of fruit against sour rot. These results suggested that M. citriensis could be used as the potential alternative of fungicides to control postharvest pathogens on citrus fruit.

Effect of Thymol and Carvacrol Encapsulated in Hp-Β-Cyclodextrin by Two Inclusion Methods against Geotrichum citri-aurantii.

Geotrichum citri-aurantii causes sour rot in citrus fruits and is responsible for important economic losses during storage. However, the availability of chemical fungicides for the control of this pathogen is limited. Thus, the aim of this research was to evaluate the antifungal efficacy of thymol and carvacrol encapsulated in 2-hydroxylpropyl-beta-cyclodextrin (HP-ß-CD) (prepared by the microwave irradiation method [MW] and solubility method [S]) for inhibition of G. citri-aurantii using in vitro bioassays broth (micro and macrodilutions methods) and inoculated food testing. Both encapsulated thymol and carvacrol were shown to be effective for inhibiting G. citri-aurantii growth in in vitro assays. Thymol was more effective in inhibiting G. citri-aurantii, while better encapsulation was provided by MW. HP-ß-CD-thymol encapsulated by MW (HP-ß-CD-thymol-MW) showed the lowest 50% effective dose (ED50 = 1.16 mM), minimum inhibitory concentration (MIC = 5.06 mM), and minimum fungicide concentration (MFC = 52.6 mM). HP-ß-CD-thymol-MW was found highly effective in reducing the growth rate and mycelial growth inhibition. Finally, HP-ß-CD-thymol-MW and HP-ß-CD-carvacrol-MW showed a higher persistent effect than thymol and carvacrol in their natural form in inhibiting this fungus. Therefore, HP-ß-CD-thymol-MW could be a promising alternative to synthetic fungicides for controlling G. citri-aurantii, the causal agent of citrus sour rot. PRACTICAL APPLICATION: Encapsulated thymol and carvacrol in HP-ß-Cyclodextrins are effective for controlling G. citri-aurantii in in vitro experiments. Encapsulation of thymol and carvacrol by microwave irradiation method (MW) was more effective than the solubility (S) method. Thymol was more effective than carvacrol, and the best results on G. citri-auriantii inhibition were achieved using the HP-ß-CD-thymol-MW method (which gave the lowest ED50 , MIC, and MFC).

Sequencing of FKS Hot Spot 1 from Saprochaete capitata To Search for a Relationship to Reduced Echinocandin Susceptibility.

Saprochaete capitata, formerly known as Geotrichum capitatum, is an emerging fungal pathogen with low susceptibility to echinocandins. Here, we report the nucleotide sequence of the S. capitata hot spot 1 region of the FKS gene (FKS HS1), which codifies for the catalytic subunit of ß-1,3-d-glucan synthase, the target of echinocandins. For that purpose, we first designed degenerated oligonucleotide primers derived from conserved flanking regions of the FKS1 HS1 segment of 12 different fungal species. Interestingly, analysis of the translated FKS HS1 sequences of 12 isolates of S. capitata revealed that all of them exhibited the same F-to-L substitution in a position that is highly related to reduced echinocandin susceptibility.

Biofilm production by Aureobasidium pullulans improves biocontrol against sour rot in citrus.

The biocontrol efficiency of Aureobasidium pullulans strain ACBL-77 against Geotrichum citri-aurantii, causal agent of sour rot in citrus, and their interactions were evaluated. For this, were evaluated the incorporation of nutrients in optimizing the antagonistic activity of the yeast, the competition for nutrients between microorganisms, the effect of nutrients on yeast cell and biofilm production and their correlation in the biocontrol of disease, the survival of yeast in citrus fruits and the interaction between microorganisms using scanning electron microscopy. Micronutrients (boric acid, cobalt chloride and ammonium molybdate) favoured the antagonistic action of A. pullulans. Ammonium sulfate 1% and sucrose 0.5% favoured the yeast during the competition between the microorganisms. The addition of ammonium sulfate (1%) in the yeast culture stimulated biofilm production and increased the antagonistic activity against the disease, as also allowed the better survival of yeast in wounded sites of citrus fruit. The yeast was found to be able to form biofilms on citrus, deforming the pathogen hyphae. These results showed the importance of the addition of nutrients in A. pullulans based-formulations when aiming for their use on a commercial scale. This is the first report of a positive correlation between the increase in the quantity of biofilm produced by A. pullulans, with increased antagonistic activity.

Characterization and Synergistic Effect of Antifungal Volatile Organic Compounds Emitted by the Geotrichum candidum PF005, an Endophytic Fungus from the Eggplant.

Plant-associated endophytes are recognized as sources of novel bioactive molecules having diverse applications. In this study, an endophytic yeast-like fungal strain was isolated from the fruit of eggplant (Solanum melongena) and identified as Geotrichum candidum through phenotypic and genotypic characterizations. This endophytic G. candidum isolate PF005 was found to emit fruity scented volatiles. The compositional profiling of volatile organic compounds (VOCs) revealed the presence of 3-methyl-1-butanol, ethyl 3-methylbutanoate, 2-phenylethanol, isopentyl acetate, naphthalene, and isobutyl acetate in significant proportion when analyzed on a time-course basis. The VOCs from G. candidum exhibited significant mycelial growth inhibition (54%) of phytopathogen Rhizoctonia solani, besides having mild antifungal activity against a few other fungi. The source of carbon as a nutrient was found to be an important factor for the enhanced biosynthesis of antifungal VOCs. The antifungal activity against phytopathogen R. solani was improved up to 91% by feeding the G. candidum with selective precursors of alcohol and ester volatiles. Furthermore, the antifungal activity of VOCs was enhanced synergistically up to 92% upon the exogenous addition of naphthalene (1.0 mg/plate). This is the first report of G. candidum as an endophyte emitting antifungal VOCs, wherein 2-penylethanol, isopentyl acetate, and naphthalene were identified as important contributors to its antifungal activity. Possible utilization of G. candidum PF005 as a mycofumigant has been discussed based upon its antifungal activity and the qualified presumption of safety status.

Use of Cistus aqueous extracts as botanical fungicides in the control of Citrus sour rot.

In this work, we investigated the in vitro and in vivo antifungal activity of aqueous extracts obtained from eight Cistus plant species against the development of Geotrichum citri-aurantii, the causal agent of citrus sour rot. The results demonstrate the in vitro effectiveness of all tested Cistus species aqueous extracts against G. citri-aurantii, the inhibition of mycelial growth ranged between 80 and 100%. Furthermore, Cistus aqueous extracts totally inhibited germination of G. citri-aurantii arthrospores at a concentration of 5 mg/mL. Among the plant species tested, C. laurifolius, C. salviifolius, C. monspeliensis, C. ladanifer and C. populifolius displayed the best fungistatic activity since the minimum inhibitory concentration (MIC) was <0.625 mg/mL. Under in vivo conditions, incidence of sour rot was lowered to 8.33% when fruits were treated with aqueous extracts of C. populifolius and C. creticus compared to 100% in the control. Disease severity was lowered to 3.74, 4.47 and 5.13% when fruits were treated with C. creticus, C. ladanifer and C. populifolius aqueous extracts, respectively. Using such these biopesticides in a replacement for synthetic fungicides or in combination with other established disease management practices could help control citrus postharvest decay in a more sustainable and eco-friendly way.

Concentrations and identification of culturable airborne fungi in underground stations of the Seoul metro.

The purpose of this study was to measure the culturable airborne fungi (CAF) concentrations in the underground subway stations of Seoul, Korea at two time points. This study measured the CAF concentrations in enclosed environments at 16 underground stations of the Seoul Metro in 2006 and 2013 and investigated the effects of various environmental factors, including the presence of platform screen doors, temperature, relative humidity, and number of passengers. CAF concentrations at the stations in 2006 were significantly higher than that at the same stations in 2013 (p < 0.001). Furthermore, there was a significant correlation between CAF concentration and relative humidity (r = 0.311, p < 0.05). Geotrichum and Penicillium were the predominant genera. The CAF concentrations in stations with an operating supply air were significantly higher than that in stations with no supply air (p < 0.001). Therefore, it is recommended that special attention be given to stations with clean supplied air to improve the indoor air quality of these subway stations.

Mold-Ripened Soft Cheeses Fortified with Date Palm Fruit Product as Functional Dairy Products.

Date fruit based products are gaining popularity among the consumers in almost all date growing countries due to its added nutritional value. Therefore, novel products were developed by combining two types of foods i.e., soft ripened cheeses and date fruit syrups or date powder. This study is the first to report the surface mold-ripened cheese production with date syrup and date powder. Model cheeses were prepared from pasteurized milk inoculated with Streptococcus thermophilus, Penicillium camemberti and Geotrichum candidum. Date syrup-1, date syrup-2, date powder or the date mixture were added at the stage of curdling. Based on the kinetic growth of the microbial groups in all the treatments, there was no change in the growth of these in various date palm product. On the contrary It may be said that addition of the date fruit product supports their growth. After 35 days, the amounts of total poly phenols were 128.3 ± 1.01, 81.8 ± 1.11, 33.5 ± 2.19, 156.23 ± 1.27 mg GAE/100 g in the cheeses support with date syrup-1, date syrup-2, date powder or the date mixture, respectively. Antioxidant activity of date fruits ranged from 80.13 IC50 (date syrup-2) to 82.23 IC50 (date syrup-1). Based on the chemical characteristics and sensory analysis, the study results showed the potential for innovative application of date products for developing new functional dairy products as an ideal medium for the delivery of biological active compounds with beneficial health effects over.

Differential gene retention as an evolutionary mechanism to generate biodiversity and adaptation in yeasts.

The evolutionary history of the characters underlying the adaptation of microorganisms to food and biotechnological uses is poorly understood. We undertook comparative genomics to investigate evolutionary relationships of the dairy yeast Geotrichum candidum within Saccharomycotina. Surprisingly, a remarkable proportion of genes showed discordant phylogenies, clustering with the filamentous fungus subphylum (Pezizomycotina), rather than the yeast subphylum (Saccharomycotina), of the Ascomycota. These genes appear not to be the result of Horizontal Gene Transfer (HGT), but to have been specifically retained by G. candidum after the filamentous fungi-yeasts split concomitant with the yeasts' genome contraction. We refer to these genes as SRAGs (Specifically Retained Ancestral Genes), having been lost by all or nearly all other yeasts, and thus contributing to the phenotypic specificity of lineages. SRAG functions include lipases consistent with a role in cheese making and novel endoglucanases associated with degradation of plant material. Similar gene retention was observed in three other distantly related yeasts representative of this ecologically diverse subphylum. The phenomenon thus appears to be widespread in the Saccharomycotina and argues that, alongside neo-functionalization following gene duplication and HGT, specific gene retention must be recognized as an important mechanism for generation of biodiversity and adaptation in yeasts.

Formation of filamentous aerobic granules: role of pH and mechanism.

Filamentous overgrowth in aerobic granular sludge processes can cause reactor failure. In this work, aerobic granules were cultivated in five identical sequencing batch reactors with acetate or glucose as the carbon source with various values of influent pH (4.5-8). Microscopic observations revealed that acidic pH, rather than the species of carbon source, epistatically controls the aerobic granules with filamentous structure. An acidic pH shifted the structure of the microbial community in the granules, such that the fungus Geotrichum fragrans was the predominant filamentous microorganism therein. The acidic pH reduced the intracellular cyclic diguanylate (c-di-GMP) content for increasing the motility of the bacteria to washout and increase the growth rate of G. fragrans on glucose or acetate, together causing overgrowth of the fungus. Maintaining the suspension under alkaline condition is proposed as an effective way to suppress filamentous overgrowth and maintain granule stability.

Characterization of mutual relations between Geotrichum candidum and Lactobacillus rhamnosus GG in milk: a quantitative approach.

The growth interactions between Geotrichum candidum and Lactobacillus rhamnosus GG were studied in milk. The effect of temperature on the growth rate of the fungus was modelled using the cardinal temperature model with inflection. The secondary modelling was applied also on the other data set containing the growth rates of G. candidum in co-culture with a commercial starter culture. The low temperature in combination with L. rhamnosus GG in co-culture showed the most negative effect on the growth rate of G. candidum. On the other hand, neither L. rhamnosus GG nor the starter culture had significant effect on the optimum and maximum temperature parameters calculated for growth of G. candidum. Their values ranged from 28.9 °C to 31.3°C and 35.3°C to 37.3°C, respectively. The quantitative data presented in the study showed a non-specific effect of lactic acid bacteria on the growth rate of G. candidum observed mainly around the optimal temperature.

Geotrichum candidum as a possible cause of bovine abortion.

Geotrichum spp. are ubiquitous, saprotrophic fungi found in soil, organic matter, and silage, as a contaminant in food products and in the digestive tracts of mammals. The current study reports a case of Geotrichum candidum infection with dermatitis in an aborted bovine fetus with skin and lung lesions. A 6-month-old aborted male Holstein Friesian fetus displayed unusual lesions on the skin of the abdomen, thorax, and head, which was excessively thickened and wrinkled. These changes corresponded to orthokeratotic hyperkeratosis, neutrophil accumulation in the stratum corneum, a pyogranulomatous inflammatory infiltrate, and superficial dermal necrosis. Moderate suppurative multifocal pneumonia was observed. Large numbers of mononuclear cells and occasional fibrin thrombi within blood vessels were found in the lungs, brain, and cerebellum. Gridley staining revealed fungal structures within the skin lesions. The mycological exam demonstrated the growth of G. candidum, and phase contrast microscopy conducted on the abomasal fluid revealed hyphae compatible with this agent. The skin lesions observed, in association with the fungus isolated, indicated that the abortion was due to G. candidum infection of the bovine fetus.

The Burkholderia contaminans MS14 ocfC gene encodes a xylosyltransferase for production of the antifungal occidiofungin.

Burkholderia contaminans strain MS14 produces the antifungal compound occidiofungin, which is responsible for significant antifungal activities against a broad range of plant and animal fungal pathogens. Occidiofungin is a cyclic glycolipopeptide made up of eight amino acids and one xylose. A 56-kb ocf gene cluster was determined to be essential for occidiofungin production. In this study, the ocfC gene, which is located downstream of ocfD and upstream of the ocfB gene in the ocf gene cluster, was examined. Antifungal activity of the ocfC gene mutant MS14KC1 was reduced against the indicator fungus Geotrichum candidum compared with that of the wild-type strain. Furthermore, the analysis of the protein sequence suggests that the ocfC gene encodes a glycosyltransferase. Biochemical analyses using nuclear magnetic resonance (NMR) and mass spectroscopy revealed that the ocfC mutant produced the occidiofungin without the xylose. The purified ocfC mutant MS14KC1 product had a level of bioactivity similar to that of the wild-type product. The revertant MS14KC1-R of the ocfC mutant produced the same antifungal activity level on plate assays and the same antifungal compound based on high-performance liquid chromatography (HPLC) and mass spectroscopy analysis as wild-type strain MS14. Collectively, the study demonstrates that the ocfC gene encodes a glycosyltransferase responsible to add a xylose to the occidiofungin molecule and that the presence of the xylose is not important for antifungal activity against Candida species. The finding provides a novel variant for future studies aimed at evaluating its use for inhibiting clinical and agricultural fungi, and the finding could also simplify the chemical synthesis of occidiofungin variants.

Inulinase production by Geotrichum candidum OC-7 using migratory locusts as a new substrate and optimization process with Taguchi DOE.

Utilization of migratory locusts (Locusta migratoria) as a main substrate due to its high protein content for inulinase (2,1-ß-d-fructan fructanohydrolase) production by Geotrichum candidum OC-7 was investigated in this study. To optimize fermentation conditions, four influential factors (locust powder (LP) concentration, sucrose concentration, pH and fermentation time) at three levels were investigated using Taguchi orthogonal array (OA) design of experiment (DOE). Inulinase yield obtained from the designed experiments with regard to Taguchi L9 OA was processed with Minitab 15 software at 'larger is better' as quality character. The results showed that optimal fermentation conditions determined as LP 30 g/l, sucrose 20 g/l, pH 6.0 and time 48 h. Maximum inulinase activity was recorded as 30.12 U/ml, which was closer to the predicted value (30.56 U/ml). To verify the results, analysis of variance test was employed. LP had the greatest contribution (71.96%) among the other factors. Sucrose had lower contribution (13.96%) than LP. This result demonstrated that LP had a strong effect on inulinase activity and can be used for enzyme production. Taguchi DOE application enhanced enzyme activity to about 3.05-fold versus unoptimized condition and 2.34-fold versus control medium. Consequently, higher inulinase production can be achieved by the utilization of an edible insect material as an alternative substrate and Taguchi DOE presents suitable optimization method for biotechnological process.