Lipolytica Cells Research Articles

Metabolic engineering of β-carotene biosynthesis in Yarrowia lipolytica.

Carotenoids, as potent antioxidant compounds, have gained extensive attention, especially in human health. In this study, the combination of CRISPR/Cas9 integration strategy and fermenter cultivation was utilized to obtain efficient β-carotene-producing Yarrowia lipolytica cell factories for potential industrial application. The introduction of the genes of Mucor circinelloides, encoding phytoene dehydrogenase (carB) and bifunctional phytoene synthase/lycopene cyclase (carRP), contributed to the heterologous production of β-carotene in Y. lipolytica XK2. Furthermore, β-carotene production was efficiently enhanced by increasing the copy numbers of the carB and carRP genes and overexpressing of GGS1, ERG13, and HMG, the genes related to the mevalonate (MVA) pathway. Thus, the optimized strain overexpressed a total of eight genes, including three copies of carRP, two copies of carB, and single copies of GGS1, HMG, and ERG13. As a consequence, strain Y. lipolytica XK19 accumulated approximately 408mg/L β-carotene in shake flask cultures, a twenty-four-fold increase compared to the parental strain Y. lipolytica XK2. 4.5g/L β-carotene was obtained in a 5-L fermenter through a combination of genetic engineering and culture optimization, suggesting a great capacity and flexibility of Y. lipolytica in the production of carotenoids.

Bacterial Cellulose-Alginate Composite Beads as Yarrowia lipolytica Cell Carriers for Lactone Production.

The demand for natural lactone gamma-decalactone (GDL) has increased in the fields of food and cosmetic products. However, low productivity during bioprocessing limits its industrial production. In this study, a novel composite porous cell carrier, bacterial cellulose-alginate (BC-ALG), was used for long-term biotransformation and production of GDL. The effects of this carrier on biotransformation and related mechanisms were investigated. BC-ALG carriers showed improved mechanical strength over ALG carriers, with their internal embedded cell pattern changed to an interconnected porous structure. In five repeated-batch biotransformation experiments, the maximum concentration of GDL obtained in culture with BC-ALG carriers was 8.37 g/L, approximately 3.7 times higher than that from the medium with an ALG carrier alone. The result indicated that multiple hydrogen bonding interactions at the interface between BC and ALG contributed to the compatibility and stability of BC-ALG carriers. On the basis of the above results, the BC-ALG composite carrier can be considered ideal for immobilisation of cells for the production of GDL on an industrial scale, and has the potential to be utilised in other biological processes.

Интенсивность свободнорадикального окисления в клетках Yarrowia lipolytica при выращивании в условиях экстремальных значений рН среды

Интенсивность свободнорадикального окисления в клетках Yarrowia lipolytica при выращивании в условиях экстремальных значений рН среды

Extraction and purification of violacein from Yarrowia lipolytica cells using aqueous solutions of surfactants

AbstractBACKGROUNDThe demand for colorants from natural bio‐based sources is increasing. Violacein is a natural purple‐blue hydrophobic pigment with interesting bioactivity whose expression in genetically modified Yarrowia lipolytica production was successfully achieved.RESULTSIn this work, several surfactants were tested in the extraction of violacein from Y. lipolytica cells, and the operational conditions were optimized to maximize the extraction yield. After the optimization, the purification of violacein using aqueous biphasic systems (ABS) composed of Tween 20 and cholinium‐based ionic liquids (ILs) was pursued. The ABS were characterized and applied in the separation of violacein from contaminant proteins, reaching a maximum selectivity of 155 with violacein being fully concentrated in the Tween 20‐rich phase, with 80% of the contaminant proteins present in the extract being removed.CONCLUSIONThis work led to a conceptual downstream process based on a first step of (solid–liquid) extraction and a second step addressing the separation of violacein from contaminant proteins using ABS. © 2019 Society of Chemical Industry

An Insight into Storage Lipid Synthesis by Yarrowia lipolytica Yeast Relating to Lipid and Sugar Substrates Metabolism.

Single cell oil (SCO) is the lipid accumulated in the cells of oleaginous microorganisms. Cellular lipids can be synthesized in two different pathways: de novo by metabolizing hydrophilic substrates and ex novo by fermenting hydrophobic substrates. The aim of the study was to evaluate the effect of carbon source (glucose and olive oil) in the culture medium on the course of microbial oil accumulation in Y. lipolytica cells. The level of selected gene expression by real time quantitative PCR method was investigated. The significant increase in expression of the POX2 gene encoding acyl-CoA oxidase II, which preferentially oxidizes long-chain acyl-CoAs formed from substrate fatty acids incorporated inside the microbial cell, was observed in medium with olive oil in relation to glucose containing medium. Noteworthily, the presence of lipid carbon substrate did not inhibit the level of ACL gene transcription coding for ATP-citrate lyase, the key enzyme of the lipid de novo accumulation process. The present study indicated that de novo lipid biosynthesis could occur despite the presence of fatty acids in the medium, and the synthesis of storage lipids in the presence of lipid carbon substrates could be carried out with the use of both pathways (de novo and ex novo).

The analysis of nanomechanical properties of Candida spp. by atomic force microscopy (AFM) method

The aim of the study was to analyze the selected nanomechanical properties of Candida spp: Candida albicans (standard strain ATCC 10231), Candida albicans (clinical strain, cultured from an oral swab), Candida lipolytica (clinical strain, cultured from a nosal swab) in atomic force microscopy (AFM). The culture Candida spp. was performed of Tryptone Soya Broth (BioMaxima). The topography and sample properties were analysed in AFM (Ntegra Spectra C from NT) and the results were carried out using NOVA 1.1.0.1824 software. C. albicans ATCC 10231 cells were significantly higher 1.81 μm (p = 0.001) from clinical strains: C. albicans (1.30 μm) and C. lipolytica (1.23 μm). C. albicans ATCC 10231 cells, and C. albicans cells of the clinical strain were softer, especially in the top parts of cells, than C. lipolytica cells. Adhesion force measured for C. albicans ATCC 10231 was 62.83 nN, and was significantly higher compared to the values obtained for C. albicans (41.93 nN, p = 0.0002 ) and C. lipolytica (41.78 nN, p = 0.0002 ). The stiffness of the Candida spp. cell surface was comparable and was in the range of 5–6 nA. The differences in height may result from different conditions in which clinical strains grow. Adhesion force can be helpful in the analysis of the degree of destruction of the cell wall by various substances. The conducted analyses showed morphological differences and the differences in mechanical properties of the researched Candida spp. This data may be important in assessing their susceptibility to the effects of various substances of a lytic nature.

An efficient method of Yarrowia lipolytica immobilization using oil- and emulsion-modified bacterial cellulose carriers

Abstract Background Yarrowia lipolytica is a nonconventional, dimorphic yeast with multiple biotechnological applications. Considering the size of Y. lipolytica cells and a plethora of its morphological forms (spherical cells or hyphae and pseudohyphae), it is highly difficult to select a suitable carrier for this useful microorganism. Bacterial cellulose (BC) is currently considered one of the most promising immobilization carriers. In the current study, the usefulness of oil- and emulsion-modified BCs as a carrier for Y. lipolytica immobilization was investigated. Static and agitated cultures were conducted in media supplemented with oil or emulsion to improve carrier porosity. Results It was found that the application of oil- and emulsion-modified BCs correlated with significantly higher efficiency of Y. lipolytica immobilization and hence higher yield than the yield achieved with an unmodified carrier. Increased efficiency of immobilization correlated with BC porosity-related parameters, which, in turn, depended on the size of oil droplets introduced into the culture medium. Moreover, changes in porosity-related parameters caused by the addition of oil or emulsion to the medium were observed when the cultures were conducted only under static conditions and not under agitated conditions. Conclusion The application of oil- and emulsion-modified BCs as carriers significantly increased the efficiency of Y. lipolytica immobilization as compared to unmodified BC. The addition of oil or emulsion to the culture medium can be a simple but effective method to modify the porosity of BC-based carriers. How to cite: Żywicka A, Wenelska K, Junka A, et al. An efficient method of Yarrowia lipolytica immobilization using oil- and emulsion-modified bacterial cellulose carrier. Electron J Biotechnol 2019;41. https://doi.org/10.1016/j.ejbt.2019.06.004 .

Genotoxic effect of caffeine in Yarrowia lipolytica cells deficient in DNA repair mechanisms.

Caffeine is a compound that can exert physiological-beneficial effects in the organism. Nevertheless, there are controversies about its protective-antioxidant and/or its negative genotoxic effect. To abound on the analysis of the possible genotoxic/antioxidant effect of caffeine, we used as research model the yeast Yarrowia lipolytica parental strain, and mutant strains (∆rad52 and ∆ku80), which are deficient in the DNA repair mechanisms. Caffeine (5mM) showed a cytostatic effect on all strains, but after 72h of incubation the parental and ∆ku80 strains were able to recover of this inhibitory effect on growth, whereas ∆rad52 was unable to recover. When cells were pre-incubated with caffeine and H2O2 or incubated with a mixture of both agents, a higher inhibitory effect on growth of mutant strains was observed and this effect was noticeably greater for the Δrad52 strain. The toxic effect of caffeine appears to be through a mechanism of DNA damage (genotoxic effect) that involves DSB generation since, in all tested conditions, the growth of Δrad52 strain (cells deficient in HR DNA repair mechanism) was more severely affected.

Immobilization pattern of morphologically different microorganisms on bacterial cellulose membranes.

The aim of this study was to assess the immobilization pattern of microorganisms characterized by varying cell shapes and sizes (rod-shaped bacteria Lactobacillus delbruecki, spherical-shaped yeast Saccharomyces cerevisiae and hyphae forms of Yarrowia lipolytica) on bacterial cellulose of various material properties. The 'adsorption-incubation' method was used for the purposes of immobilization. The immobilization pattern included adsorption efficiency, ability of the immobilized cells to multiply within the carrier expressed as incubation efficiency and the degree of release of the immobilized cells from the carrier. The efficiency of adsorption and incubation was affected by the morphology of the immobilized cells and increased together with cellulose surface area. For smaller bacterial cells a higher level of loading was obtained on the same surface as compared to larger yeast cells. During incubation, the number of immobilized bacterial and yeast cells increased significantly in comparison to the number of cells adsorbed on the carrier during the adsorption step. Despite the morphological differences between the S. cerevisiae and Y. lipolytica cells, there were no statistically significant differences in the efficiency of adsorption and incubation. It was also revealed that the release ratio values obtained for L. delbruecki and S. cerevisiae increased along with cellulose surface area. Interestingly, Y. lipolytica cells in the pseudohyphae and hyphae forms penetrated deeply into the three-dimensional network of BC nanofibrils which prevented subsequent cell release. It was confirmed that carrier selection must be individually matched to the type of immobilized cells based especially on its porosity-related parameters.

Use of Yarrowia lipolytica Lipase Immobilized in Cell Debris for the Production of Lipolyzed Milk Fat (LMF).

Lipase immobilized on Yarrowia lipolytica cell debris after sonication of yeast cells (LipImDebri) was used in hydrolysis reaction as a novel strategy to produce lipolyzed milk fat (LMF). Extracellular (4732.1 U/L), intracellular (130.0 U/g), and cell debris (181.0 U/g) lipases were obtained in a 4 L bioreactor using residual frying oil as inducer in 24 h fermentation process. LipImDebri showed a good operational stability retaining 70% of lipolytic activity after the second cycle and 40% after the fourth. The highest degree of hydrolysis (28%) was obtained with 500 mg LipImDebri for 6 h of lipolysis of anhydrous milk fat. LMF produced with LipImDebri presented high contents of oleic (35.2%), palmitic (25.0%), and stearic (15.4%) acids and considerable amounts of odor-active short and medium chain fatty acids (C:4–C:10) (8.13%).

Peptide stabilized gold and silver nanoparticles derived from the mangrove isolate Pseudoalteromonas lipolytica mediate dye decolorization

A mangrove associated halotolerant bacterium identified as Pseudoalteromonas lipolytica mediated the synthesis of gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs). Interaction of P. lipolytica cells with chloroauric acid (HAuCl4) resulted in the synthesis of cell associated AuNPs that could be released in an extracellular manner after sonication. For obtaining extracellular AgNPs, pre-grown cells were subjected to osmotic shock treatment by re-suspending cells in distilled water. The preparation was centrifuged and the filtered supernatant was challenged with 4 mM AgNO3. The biogenic NPs were characterized on the basis of UV–vis spectroscopy, Field Emission Scanning Electron Microscope observations, Transmission Electron Microscope studies, X-Ray Diffraction profiles and Energy-Dispersive X-ray spectroscopy. The size of AuNPs and AgNPs ranged between 10–30 nm and 5–15 nm, respectively. The active components in the preparation after osmotic shock treatment were subjected to thin layer chromatography, Fourier Transform Infrared spectroscopy and liquid chromatography mass spectrometry (LCMS) analysis. Five oligopeptides, Ile-Arg, His-Asp-Asp, Glu-Ile-Lys, Lys-Asp-Asn, and Arg-Arg-Gln were identified in the active band. AuNPs brought about 91% decolorization of methylene blue and congo red dyes. With AgNPs, 92% of congo red was decolorized. The rate constants for methylene blue and congo red decolorization using AuNPs were 0.2545 and 0.1587 min−1, respectively. For congo red dye decolorization by AgNPs the rate constant was 0.1882 min−1. These peptide stabilized NPs could be used as effective catalysts for sodium borohydride mediated decolorization of methylene blue and congo red dyes.

Utilização de leveduras para a produção de biomassa em extrato de palma forrageira (Opuntia ficus-indica)

Opuntia ficus-indica é uma cactácea utilizada na produção de bioprodutos, sendo rica em carboidratos e reconhecida como fonte alternativa de alimento para agricultura de subsistência. Este trabalho objetivou avaliar a utilização do extrato líquido de O. ficus-indica como substrato para o crescimento de leveduras de interesse biotecnológico. Candida utilis, C. lipolytica, Saccharomyces cerevisiae e Kluyveromyces marxianus foram cultivadas no extrato obtido após tratamento da cactácea. Alíquotas do cultivo microbiano foram coletadas para análise de biomassa, açúcares redutores, pH e parâmetros cinéticos. K. marxianus apresentou maior taxa de crescimento (µmax= 0,25h-1) e maior porcentagem de consumo de açúcar (95,68%). C. utilis obteve melhor rendimento de biomassa em relação ao substrato consumido (Yx/s=2,69). C. lipolytica se destacou por exibir melhor perfil cinético uma vez que apresentou maiores valores de produção de biomassa (∆Xmax=10,93 g/L) e produtividade (P= 0,58 g/L.h-1) em 24 horas de cultivo. A influência de diferentes concentrações salinas no crescimento de C. lipolytica cultivada em extrato de O. ficus-indica foi testada com NaCl e KCl (0,1M, 0,25M, 0,50M, 1M, 2M e 4M). No crescimento sob estresse a levedura foi tolerante às concentrações salinas (até 1M de NaCl e a 0,5M para KCl). As concentrações maiores inibiram o crescimento celular. A concentração de 4M foi considerada letal para ambas as células leveduriformes e a 2M sub-letal. As células de C. lipolytica expressaram adaptação contra estresse osmótico até a concentração de 1M. Os resultados sugerem a utilização desta cactácea como substrato alternativo para a produção de biomassa microbiana.

Carbohydrate Spectrum of Extremophilic Yeasts Yarrowia lipolytica under pH Stress

Alterations in the concentrations of cell cytosol carbohydrates of polyextremophilic yeasts Yarrowia lipolytica under stresses of diverse nature were observed. Under pH stress, mannitol was the main storage carbohydrate (up to 89% of the total cytosol carbohydrates), while arabitol, glucose, and inositol were present in insignificant amounts (3 to 6%). Experiments with inhibition of de novo mannitol synthesis by bis(p-nitrophenyl) disulfide revealed that the cytoprotective effect of mannitol was most noticeable in the cells grown under acidic conditions (pH 4.0), while the role of catalase and superoxide dismutase, the enzymes of the first line of antioxidant protection, increased under alkaline conditions (pH 9.0). The constitutively high mannitol content in Y. lipolytica cells was hypothesized to be a part of the core mechanism of stress resistance in this yeast species.

Phenol Is the Initial Product Formed during Growth and Degradation of Bromobenzene by Tropical Marine Yeast, Yarrowia lipolytica NCIM 3589 via an Early Dehalogenation Step.

Bromobenzene (BrB), a hydrophobic, recalcitrant organic compound, is listed by the environmental protection agencies as an environmental and marine pollutant having hepatotoxic, mutagenic, teratogenic, and carcinogenic effects. The tropical marine yeast Yarrowia lipolytica 3589 was seen to grow aerobically on BrB and displayed a maximum growth rate (μmax) of 0.04 h-1. Furthermore, we also observed an increase in cell size and sedimentation velocity for the cells grown on BrB as compared to the glucose grown cells. The cells attached to the hydrophobic bromobenzene droplets through its hydrophobic and acid–base interactions. The BrB (0.5%, 47.6 mM) was utilized by the cells with the release of a corresponding amount of bromide (12.87 mM) and yielded a cell mass of 1.86 g/L after showing 34% degradation in 96 h. Maximum dehalogenase activity of 16.16 U/mL was seen in the cell free supernatant after 24 h of growth. Identification of metabolites formed as a result of BrB degradation, namely, phenol, catechol, cis, cis muconic acid, and carbon dioxide were determined by LC–MS and GC–MS. The initial attack on bromobenzene by Y. lipolytica cells lead to the transient accumulation of phenol as an early intermediate which is being reported for the first time. Degradation of phenol led to catechol which was degraded by the ortho- cleavage pathway forming cis, cis muconic acid and then to Krebs cycle intermediates eventually leading to CO2 production. The study shows that dehalogenation via an extracellular dehalogenase occurs prior to ring cleavage with phenol as the preliminary degradative compound being produced. The yeast was also able to grow on the degradative products, i.e., phenol and catechol, to varying degrees which would be of potential relevance in the degradation and remediation of xenobiotic environmental bromoaromatic pollutants such as bromobenzene.

New yeast models for studying mitochondrial morphology as affected by oxidative stress and other factors

The overwhelming majority of investigations on mitochondrial morphology were performed using S. cerevisiae. In this study we showed the benefits of applying new model organisms including petite-negative D. magnusii and Y. lipolytica yeasts for visualization of mitochondrial fragmentation. Normally giant D. magnusii cells and filament-like Y. lipolytica cells contain the highly structured mitochondrial reticulum. Oxidative stress mediated by tert-butyl hydroperoxide triggered mitochondrial fragmentation in yeasts. In D. magnusii mitochondrial fragmentation was also induced by impairing the oxidative phosphorylation system. Higher prooxidant concentrations caused cell death. Cationic lipophilic antioxidant SkQ1 acted downstream of the excessive ROS production and prevented partially or almost totally oxidative stress and related mitochondrial fragmentation and cell death. We believe that utility of D. magnusii and Y. lipolytica yeasts as a “living test tube” would be useful for providing new information concerning the interplay between mitochondrial dynamics and mitochondrial dysfunction, cell cycle, aging, mitophagy and cell death.

Iterative integration of multiple-copy pathway genes in Yarrowia lipolytica for heterologous β-carotene production

β-Carotene is a terpenoid molecule with high hydrophobicity that is often used as an additive in foods and feed. Previous work has demonstrated the heterologous biosynthesis of β-carotene from an intrinsic high flux of acetyl-CoA in 12 steps through 11 genes in Yarrowia lipolytica. Here, an efficient biosynthetic pathway capable of producing 100-fold more β-carotene than the baseline construct was generated using strong promoters and multiple gene copies for each of the 12 steps. Using fed-batch fermentation with an optimized medium, the engineered pathway could produce 4g/L β-carotene, which was stored in lipid droplets within engineered Y. lipolytica cells. Expansion of these cells for squalene production also demonstrated that Y. lipolytica could be an industrially relevant platform for hydrophobic terpenoid production.

Mitochondrial cytopathies: Their causes and correction pathways

Mitochondrial cytopathies are a heterogeneous group of systemic disorders caused by mutations in mitochondrial or nuclear genome. The review presents some data on pathogenic mutations in mitochondrial DNA leading to the imbalance in the oxidation phosphorylation processes and energy metabolism in the cells and eventually to the development of mitochondrial cytopathy. The pathways of medicated correction are examined, which are aimed at obtaining optimal energy efficiency of mitochondria with impaired functions, increase of the efficiency of energy metabolism in the tissues, as well as prevention of mitochondrial membrane damage by free radicals using antioxidants and membrane protectors. A conclusion is drawn on the inefficiency of currently used therapeutic strategies and the necessity of new approaches, which can be gene therapy of mitochondrial diseases. Some modern methods for gene defects correction, capable of restoring or removing the damaged gene, expressing full gene product, or blocking the mutant or strange genes work are analyzed. It is shown that the described approaches to the gene therapy of human mitochondrial diseases demand the introduction of foreign sequences into nuclear or mitochondrial genome of a living person, which completely excludes their practical application because of the uncertainty of the outcome. A perspective approach in solving this problem may be a creation of a system allowing the correction of defect genes without introducing synthetic nucleotides into the human genome. Phenotypic selection combined with a capacity of homologous recombination, artificially imparted to mitochondria of yeast Yarrowia lipolytica, allows for replication of intact human mitochondrial DNA in yeast mitochondria, supporting a full-size native human mitochondrial DNA in the yeast cells and eliminating pathogenic mutations by means of standard sitedirected PCR mutagenesis. After the correction in the Y. lipolytica cells, copies of mitochondrial DNA of an individual patient may be returned to him using the transfection of mesenchymal stromal cells followed by selection of transfectants grown in minimal culture media, in which the cells with higher respiratory mitochondrial activity will gain the advantage.

Construction of recombinant Yarrowia lipolytica and its application in bio-transformation of lignocellulose

ABSTRACT Lignocellulose is a polysaccharide and an abundant biomass resource that widely exists in grains, beans, rice, and their by-products. Over 10 million tons of lignocellulose resources and processing products are produced every year in China. Three recombinant Y. lipolytica strains with cellulase (β-glucosidase, endoglucanase and cellobiohydrolase) were constructed. The enzymatic activities of these enzymes were 14.181 U/mL, 16.307 U/mL, and 17.391 U/mL, respectively. The whole cell cellulases were used for a stover bio-transformation. The celluloses in the stover were partly degraded by the cellulases, and the degradation products were transformed into single cell protein (SCP) by the Y. lipolytica cells. After 15 d of fermentation with the whole cell cellulases, the protein content of the maize stover and the rice straw reached 16.23% and 14.75%, which increased by 168.26% and 161.52% compared with the control, respectively. This study provides a new stage for the efficient utilization of stover in the feed industry.

A molecular genetic toolbox for Yarrowia lipolytica

BackgroundYarrowia lipolytica is an ascomycete yeast used in biotechnological research for its abilities to secrete high concentrations of proteins and accumulate lipids. Genetic tools have been made in a variety of backgrounds with varying similarity to a comprehensively sequenced strain.ResultsWe have developed a set of genetic and molecular tools in order to expand capabilities of Y. lipolytica for both biological research and industrial bioengineering applications. In this work, we generated a set of isogenic auxotrophic strains with decreased non-homologous end joining for targeted DNA incorporation. Genome sequencing, assembly, and annotation of this genetic background uncovers previously unidentified genes in Y. lipolytica. To complement these strains, we constructed plasmids with Y. lipolytica-optimized superfolder GFP for targeted overexpression and fluorescent tagging. We used these tools to build the “Yarrowia lipolytica Cell Atlas,” a collection of strains with endogenous fluorescently tagged organelles in the same genetic background, in order to define organelle morphology in live cells.ConclusionsThese molecular and isogenetic tools are useful for live assessment of organelle-specific protein expression, and for localization of lipid biosynthetic enzymes or other proteins in Y. lipolytica. This work provides the Yarrowia community with tools for cell biology and metabolism research in Y. lipolytica for further development of biofuels and natural products.

Enhanced production of insect raw-starch-digesting alpha-amylase accompanied by high erythritol synthesis in recombinant Yarrowia lipolytica fed-batch cultures at high-cell-densities

Abstract Recently we reported on raw-starch-digesting ability of alpha-amylase from an insect Sitophilus oryzae (SoAMY) expressed in recombinant Yarrowia lipolytica cells, and demonstrated its usefulness in simultaneous saccharification and fermentation processes with industrial yeasts. In this study we applied fed-batch cultures of Y. lipolytica 4.29 strain reaching high-cell-densities (up to 70 [gDCW/L]), to enhance SoAMY production. SoAMY activity in the medium reached the peak value of 22,979.23 ± 184 [AU/L], at volumetric productivity of 121.58 ± 1.75 [AU/L/h], and yield of 71.83 ± 3.08 [AU/gglycerol], constituting roughly 160-fold improvement, compared to the best previous result. The cultivations were accompanied by high production of erythritol (83.58 [g/L]), at the marginal production of mannitol (5.46 [g/L]). Elementary analyses of media constituents, the enzyme and the yeast biomass gave better insight into carbon and nitrogen fluxes distribution. Due to application of genetic engineering and bioprocess engineering strategies, the insect-derived enzyme can be produced at the quantities competitive to microbial catalysts.