Growth and by-product profiles of Kluyveromyces marxianus cells immobilized in foamed alginate.

The aim of this research was to study how the yeast cell immobilization technique influences the growth and fermentation profiles of Kluyveromyces marxianus cultivated on apple/chokeberry and apple/cranberry pomaces. Encapsulation of the cells was performed by droplet formation from a foamed alginate solution. The growth and metabolic profiles were evaluated for both free and immobilized cells. Culture media with fruit waste produced good growth of free as well as immobilized yeast cells. The fermentation profiles of K. marxianus were different with each waste material. The most varied aroma profiles were noted for immobilized yeast cultivated on apple/chokeberry pomace.

Influence of carbon and nitrogen source on production of volatile fragrance and flavour metabolites by the yeast Kluyveromyces marxianus.

The yeast Kluyveromyces marxianus produces a range of volatile molecules with applications as fragrances or flavours. The purpose of this study was to establish how nutritional conditions influence the production of these metabolites. Four strains were grown on synthetic media, using a variety of carbon and nitrogen sources and volatile metabolites analysed using gas chromatography-mass spectrometry (GC-MS). The nitrogen source had pronounced effects on metabolite production: levels of the fusel alcohols 2-phenylethanol and isoamyl alcohol were highest when yeast extract was the nitrogen source, and ammonium had a strong repressing effect on production of 2-phenylethyl acetate. In contrast, the nitrogen source did not affect production of isoamyl acetate or ethyl acetate, indicating that more than one alcohol acetyl transferase activity is present in K. marxianus. Production of all acetate esters was low when cells were growing on lactose (as opposed to glucose or fructose), with a lower intracellular pool of acetyl CoA being one explanation for this observation. Bioinformatic and phylogenetic analysis of the known yeast alcohol acetyl transferases ATF1 and ATF2 suggests that the ancestral protein Atf2p may not be involved in synthesis of volatile acetate esters in K. marxianus, and raises interesting questions as to what other genes encode this activity in non-Saccharomyces yeasts. Identification of all the genes involved in ester synthesis will be important for development of the K. marxianus platform for flavour and fragrance production.

Bioflavour production from tomato and pepper pomaces by Kluyveromyces marxianus and Debaryomyces hansenii.

Bioflavours are called natural flavour and/or fragrance compounds which are produced using metabolic pathway of the microorganism and/or plant cells or their enzyme systems with bioengineering approaches. The aim of this study was to investigate bioflavour production from tomato and red pepper pomaces by Kluyveromyces marxianus and Debaryomyces hansenii. Obtained specific growth rates of K. marxianus and D. hansenii in tomato pomace were 0.081/h and 0.177/h, respectively. The bioflavour profile differed between the yeasts. Both yeasts can produce esters and alcohols such as phenyl ethyl alcohol, isoamyl alcohol, isoamyl acetate, phenyl ethyl acetate and isovaleric acid. "Tarhana" and "rose" were descriptive flavour terms for tomato and pepper pomaces fermented by K. marxianus, respectively. Tomato pomace fermented by D. hansenii had the most intense "green bean" flavour while "fermented vegetable" and "storage/yeast" were defined as characteristic flavour terms for pepper pomaces fermented by D. hansenii.

Effect of emulsifiers on oil separation problem and quality characteristics of Tahin Helva during storage.

Tahin helva is one of Turkish traditional confectionary foods. Sesame paste, sugar syrup and soapwort extract were used for tahin helva's production. The purpose of this study was to solve the oil separation problem occurred in tahin helva by using food additives permitted as an emulsifier for tahin helva in the Turkish and EU Codex. Sorbitan tristearate (STS), sorbitan monopalmitate (SMP) and their combinations (STS:SMP, 1:1) were added to tahin helva, then helva samples were stored at the storage temperatures of 20 °C, 30 °C, and 40 °C. The emulsifiers did not have any effect on physical, chemical and sensory properties of tahin helva except on its oil content, whereas storage temperature and storage time had effects on those properties of helva. STS:SMP combination was found to be statistically more effective than SMP and STS formulations in preventing oil separation. Penetration force of tahin helva has increased by increasing the storage temperature and storage time. It was found that there was a decrease in -a colour value of helva through the storage time at 20 °C and 30 °C, while there was an increase in that value at 40 °C. The highest change in the moisture content of all helva samples through storage was observed at the storage temperature of 20 °C. Sensory scores for appearance, texture and flavour characteristics of helva have decreased significantly during the storage period.

Optimizing bioreactor conditions for Spirulina fermentation by Lactobacillus helveticus and Kluyveromyces marxianus: Impact on chemical & bioactive properties.

This study focused on optimizing the production of fermented Spirulina (FS) products using a bioactivity-guided strategy with Lactobacillus helveticus B-4526 and Kluyveromyces marxianus Y-329 in a 3-L bioreactor. Various operating conditions, including aeration rates and pH modes, were tested. While both microorganisms thrived under all conditions, the "cascade" mode, controlling dissolved oxygen, enhanced protein hydrolysis and antioxidant activity, as confirmed by SDS-PAGE and DPPH/TEAC assays, respectively. Screening revealed that "cascade" FS significantly decreased viability of colon cancer cells (HT-29) in a dose-dependent manner, with up to a 72 % reduction. Doses ≤ 500 µg mL-1 of "cascade" FS proved safe and effective in suppressing NO release without compromising cellular viability. Additionally, "cascade" FS exhibited diverse volatile organic compounds and reducing the characteristic "seaweed" aroma. These findings highlight "cascade" FS as a promising alternative food source with improved bioactive properties, urging further exploration of its bioactive compounds, particularly bioactive peptides.

Influence of Different Bacteria Species in Chemical Composition and Sensory Properties of Fermented Spirulina.

Fermented Spirulina (FS) could be a good strategic approach for diversifying algae-derived formulations to a current functional food market. In this study, microbial properties, total and free amino acids, protein hydrolysis, volatile organic compounds (VOCs) and sensory properties of FS and unfermented Spirulina (unFS) products by four different Lactic acid bacteria (LAB) and three different Bacillus strains were examined. The highest proteolytic activity with LAB strains was confirmed by SDS-PAGE. The increase in both total amino acid (TAA) and total essential amino acid (TEAA) concentrations were in the highest level for FS products by Bacillus strains, 70535.5 µM and 22295.4 µM, respectively. The pyrazine content, the most prevalent VOCs in unFS, reduced more remarkably in FS by LAB strains. Furthermore, the most acceptable sensory characteristics were obtained with FS products by LAB strains. These findings will provide insights toward achieving the industrialization of FS products.

Production of flavor compounds from rice bran by yeasts metabolisms of Kluyveromyces marxianus and Debaryomyces hansenii.

The aim of this study was to evaluate the biosynthesis of flavor compounds from rice bran by fermentation facilitated by Kluyveromyces marxianus and Debaryomyces hansenii. The growth of both yeasts was assessed by specific growth rates and doubling time. The biosynthesis of flavor compounds was evaluated by gas chromatography-olfactometry (GC-O), gas chromatography-mass spectrometry (GC-MS), and Spectrum™ sensory analysis. The specific growth rate (µ) and doubling time (td) of K. marxianus was calculated as 0.16/h and 4.21h, respectively, whereas that of D. hansenii was determined as 0.13/h and 5.33h, respectively. K. marxianus and D. hansenii produced significant levels of higher alcohols and acetate esters from rice bran. Results showed that K. marxianus can produce 827.27 µg/kg of isoamyl alcohol, 169.77 µg/kg of phenyl ethyl alcohol, and 216.08 µg/kg of phenyl ethyl acetate after 24-h batch fermentation. A significant amount of isovaleric acid was also synthesized by K. marxianus (4013 µg/kg) after the batch fermentation of 96 h. 415.64 µg/kg of isoamyl alcohol and 135.77 µg/kg of phenyl ethyl acetate was determined in rice bran fermented by D. hansenii after 24-h fermentation. Fermented cereals and rose were the characteristic flavor descriptors of the fermented rice bran samples. Rose flavor in fermented rice bran samples was found to be associated with phenyl ethyl alcohol, phenyl ethyl acetate, isoamyl acetate, and guaiacol. Thus, the findings of this study demonstrate that the valorization of rice bran can be achieved with the production of natural flavor compounds by yeast metabolism.

Biosynthesis of eight-carbon volatiles from tomato and pepper pomaces by fungi: Trichoderma atroviride and Aspergillus sojae.

The aim of this study was to investigate the possibility of using tomato and red pepper pomaces for the production eight-carbon volatiles by Trichoderma atroviride and Aspergillus sojae. The fermentation of tomato and pepper pomace-based media by both moulds was conducted in shake flasks and bioreactors. Microbial growth behaviours and fermentation abilities of T. atroviride and A. sojae under both fermentation conditions were followed by microbial counting. The production of flavours from tomato and pepper pomaces by fungal metabolism was determined by gas chromatography-olfactometry, gas chromatography-mass spectrometry and sensory analysis. The results showed that T. atroviride grew faster than A. sojae, and the survival of T. atroviride in the tomato pomace was longer than that of A. sojae. However, T. atroviride grew slower than A. sojae in the pepper pomace. Eight-carbon flavour compounds, including (Z)-1,5-octadien-3-ol, 1-octen-3-ol, (E)-2-octenal and (E)-2-octenol, were produced by T. atroviride and A. sojae from the tomato and pepper pomaces. The highest production levels (265.55 ± 2.79 and 187.47 ± 0.92 µg kg-1) were observed for 1-octen-3-ol in the tomato fermentation by T. atroviride and A. sojae, respectively. The relationships between volatile compounds and their flavour characteristics in tomato and pepper pomaces were analysed using principal component analysis.

Production of flavor compounds from olive mill waste by Rhizopus oryzae and Candida tropicalis.

The purpose of this study was to investigate the production of flavor compounds from olive mill waste by microbial fermentation of Rhizopus oryzae and Candida tropicalis. Olive mill waste fermentations were performed in shake and bioreactor cultures. Production of flavor compounds from olive mill waste was followed by Gas Chromatography-Mass spectrometry, Gas chromatography- olfactometry and Spectrum Sensory Analysis®. As a result, 1.73-log and 3.23-log cfu/mL increases were observed in the microbial populations of R. oryzae and C. tropicalis during shake cultures, respectively. C. tropicalis can produce a higher concentration of d-limonene from olive mill waste than R. oryzae in shake cultures. The concentration of d-limonene was determined as 185.56 and 249.54µg/kg in the fermented olive mill waste by R. oryzae and C. tropicalis in shake cultures respectively. In contrast, R. oryzae can produce a higher concentration of d-limonene (87.73µg/kg) d-limonene than C. tropicalis (11.95µg/kg) in bioreactor cultures. Based on sensory analysis, unripe olive, wet towel, sweet aromatic, fermented aromas were determined at high intensity in olive mill waste fermented with R. oryzae meanwhile olive mill waste fermented with C. tropicalis had only a high intensity of unripe olive and oily aroma.

Pigment and color stability of beetroot betalains in cow milk during thermal treatment.

Thermal stability of beetroot betalains in cow milk was determined during heating at 70-90 °C. Changes in color values of colored milk were also investigated. Degradation of betalains followed first order kinetics while changes of L(∗), Hue angle and Chroma values fitted zero order and first order kinetic, respectively. Reaction rate for degradation of betalains, L(∗), Hue angle and Chroma values ranged between 1.588-30.975 × 10(-3) min(-1), 90.50-379.75 L(∗)min(-1), 0.581-5.008 Hue anglemin(-1) and 3.250-19.750 × 10(-3) min(-1), respectively. Between 70 and 90 °C, activation energy for the degradation of betalains was 42.449 kJ mol(-1). L(∗) values was more stable than Hue angle and Chroma color values in colored milk during heating. 74.150 kJ mol(-1), 111.174 kJ mol(-1) and 93.311 kJ mol(-1) of activation energy values were found for L(∗), Hue angle and Chroma values of milk, respectively. Significant positive and negative linear correlations were determined between betalains and color values. Multiple regression models were also established to predict the content of betalains in milk during thermal process by using color values.

Volatile metabolites produced from agro-industrial wastes by Na-alginate entrapped Kluyveromyces marxianus.

The aim of this study was to evaluate the effects of alginate entrapment on fermentation metabolites of Kluyveromyces marxianus grown in agrowastes that served as the liquid culture media. K. marxianus cells entrapped in Na-alginate were prepared using the traditional liquid-droplet-forming method. Whey and pomaces from processed tomatoes, peppers, and grapes were used as the culture media. The changes in the concentrations of sugar, alcohol, organic acids, and flavor compounds were analyzed using gas chromatography-mass spectrometry (GC-MS) and high pressure liquid chromatography (HPLC). Both free and entrapped, K. marxianus were used individually to metabolize sugars, organic acids, alcohols, and flavor compounds in the tomato, pepper, grape, and acid whey based media. Marked changes in the fermentation behaviors of entrapped and free K. marxianus were observed in each culture. A 1.45-log increase was observed in the cell numbers of free K. marxianus during fermentation. On the contrary, the cell numbers of entrapped K. marxianus remained the same. Both free and entrapped K. marxianus brought about the fermentation of sugars such as glucose, fructose, and lactose in the agrowaste cultures. The highest volume of ethanol was produced by K. marxianus in the whey based media. The concentrations of flavor compounds such as ethyl acetate, isoamyl alcohol, isoamyl acetate, 2-phenylethyl isobutyrate, phenylethyl acetate, and phenylethyl alcohol were higher in fermented agrowaste based media compared to the control.

Investigation of antimicrobial activity and entA and entB genes in Enterococcus faecium and Enterococcus faecalis strains isolated from naturally fermented Turkish white cheeses.

In this research, the antagonistic effects of Enterococcus faecalis and E. faecium bacteria isolated from naturally fermented Turkish white cheeses, produced without starter culture, against Listeria monocytogenes, L. innocua, L. ivanovii, Staphylococcus aureus, and E. faecalis were evaluated. The presence of entA and entB genes was also detected in the isolates, which had antimicrobial activity. Total 71 strains of E. faecalis and 7 strains of E. faecium were tested; 20 of E. faecalis and none of E. faecium strains showed antimicrobial activity against the tested bacteria using agar spot method. Among E. faecalis strains, which had antimicrobial activity, three strains contained both entA and entB genes, two strains carried only entA gene, and five strains had only entB gene. These cheese-sourced enterococcal strains or their enterocins should be considered to be used for food preservation especially in the dairy industry.

Production of flavor compounds from olive mill waste by Rhizopus oryzae and Candida tropicalis

Abstract The purpose of this study was to investigate the production of flavor compounds from olive mill waste by microbial fermentation of Rhizopus oryzae and Candida tropicalis. Olive mill waste fermentations were performed in shake and bioreactor cultures. Production of flavor compounds from olive mill waste was followed by Gas Chromatography–Mass spectrometry, Gas chromatography- olfactometry and Spectrum Sensory Analysis ®. As a result, 1.73-log and 3.23-log cfu/mL increases were observed in the microbial populations of R. oryzae and C. tropicalis during shake cultures, respectively. C. tropicalis can produce a higher concentration of d-limonene from olive mill waste than R. oryzae in shake cultures. The concentration of d-limonene was determined as 185.56 and 249.54 µg/kg in the fermented olive mill waste by R. oryzae and C. tropicalis in shake cultures respectively. In contrast, R. oryzae can produce a higher concentration of d-limonene (87.73 µg/kg) d-limonene than C. tropicalis (11.95 µg/kg) in bioreactor cultures. Based on sensory analysis, unripe olive, wet towel, sweet aromatic, fermented aromas were determined at high intensity in olive mill waste fermented with R. oryzae meanwhile olive mill waste fermented with C. tropicalis had only a high intensity of unripe olive and oily aroma.