Scale-up of extremely low temperature fermentations of grape must by wheat supported yeast cells

doi:10.1016/j.biortech.2010.04.031

Bioresource Technology

Volume 101, Issue 19, October 2010, Pages 7484-7491

https://doi.org/10.1016/j.biortech.2010.04.031 Get rights and content

Abstract

A new biocatalyst was prepared by immobilization of Saccharomyces cerevisiae AXAZ-1 yeast cells on whole wheat grains. This biocatalyst was used for 30 repeated batch fermentations of glucose and grape must at various temperatures. The biocatalyst retained its operational stability for a long period and it was proved capable to produce dry wines of fine clarity even at extremely low temperatures (5 °C). After the completion of these fermentations the new biocatalyst was used in a scale-up system of 80 L for wine making at ambient (20 °C) and extremely low temperatures (2 °C). The scale-up process did not affect the fermentative ability of biocatalyst, even at low temperatures, while the produced wines had almost the same improved aromatic profile compare to free cells as revealed by GC and GC–MS analyses. More specifically the results showed that both systems with immobilized cells (laboratory scale and 80 L bioreactor) increased the formation of esters and produced wines with improved aromatic profile compared to those with free cells. Finally an increase in the percentages of total esters and a decrease in those of higher alcohols was observed in lower fermentation temperatures.

Introduction

In recent years several immobilized cell systems have been proposed for use in alcoholic fermentation, due to the several technical and economical advantages compared to free cells systems (Kourkoutas et al., 2004). However, full industrial use is limited to the production of sparkling wines (Fumi et al., 1988, Colagrande et al., 1994). The supports that are used for immobilization in wine making must be of food grade purity, low cost, abundant in nature, easy to handle, suitable for low temperature fermentations and improve wine quality. In recent years supports of food grade purity, such as gluten pellets (Iconomopoulou et al., 2002), brewer’s spent grains (Kopsahelis et al., 2007), sugarcane (Chandel et al., 2009), delignified cellulosic material (Kourkoutas et al., 2002), potatoes (Kandylis and Koutinas, 2008) and corn starch gel (Kandylis et al., 2008), have been proposed as ideal for yeast immobilization for wine making and alcohol production. The use of alcohol resistant and cryotolerant yeasts immobilized on these supports led to low temperature fermentations producing wines with excellent taste and aroma.

The use of immobilized cells on starchy supports in wine making, especially at low temperatures, led to wines with improved taste and aroma, while the reduced activation energy and the higher reaction rate constant in the case of immobilized cells led to the conclusion that these supports may behave as catalysts or promoters of the enzymes involved in the process (Kandylis and Koutinas, 2008, Kandylis et al., 2008).

Among the major cereals, wheat is the staple food in the diets of a large segment of the world’s population. It accounts nearly 20–80% of the total food consumption in various regions of the world. Wheat is produced worldwide in large amounts and can be used as a support for cell immobilization reducing the cost compared with gluten pellets and starch that need a costly process for isolation. Likewise, wheat does not need the delignification cost that is necessary in the case of brewer spent grains.

Therefore the aim of the present study was to evaluate the use of wheat as a support for yeast immobilization suitable for ambient and low temperature fermentations and its scale-up from laboratory to semi-industrial scale.

Section snippets

Yeast strain and media

The alcohol resistant and cryotolerant Saccharomyces cerevisiae AXAZ-1, isolated from Greek vineyard plantation was used in the present study (Kopsahelis et al., 2009). It was grown on culture medium consisting of 4 g yeast extract/L, 1 g (NH₄)₂SO₄/L, 1 g KH₂PO₄/L, 5 g MgSO₄ × 7H₂O/L, and 40 g glucose monohydrate/L at 30 °C and centrifuged at 5000 rpm for 10 min. All media were sterilized at 130 °C and 1.5 atm for 15 min. Must of roditis grape variety, was provided from the local wine industry “Achaia

Results and discussion

It is the first time that whole wheat grains were evaluated as supports for yeast cell immobilization and their feasibility for repeated batch fermentation of glucose and grape must at various temperatures was examined. Whole wheat grains were boiled for 15 min and after sterilization and cooling, immobilization took place by mixing the wheat grains with a liquid culture of yeast cells and the system left to ferment for ∼8 h. The immobilization was confirmed by electron microscopy, showing yeast

Conclusions

In the present study wheat grains supported biocatalyst proved capable for wine making in a wide range of temperatures in laboratory scale and in a scale-up system of 80 L, producing quality wines with great aroma. The growth and immobilization of cells and fermentations were carried out at the same bioreactor leading to a reduction of investment and operational cost. Thus, industrialization of the propose system has a great potential, considering the high fermentation efficiency and high

Acknowledgement

This work is part of the 03ED657 research project, implemented within the framework of the “Reinforcement Programme of Human Research Manpower” (PENED) and co-financed by National and Community Funds (20% from the Greek Ministry of Development-General Secretariat of Research and Technology and 80% from EU-European Social Fund).

References (30)

A.K. Chandel et al.
Use of Saccharum spontaneum (wild sugarcane) as biomaterial for cell immobilization and modulated ethanol production by thermotolerant Saccharomyces cerevisiae VS₃
Bioresour. Technol.
(2009)
M. Iconomopoulou et al.
Low temperature and ambient temperature wine making using freeze dried immobilized cells on gluten pellets
Process Biochem.
(2002)
N. Kopsahelis et al.
Comparative study of spent grains and delignified spent grains as yeast supports for alcohol production from molasses
Bioresour. Technol.
(2007)
N. Kopsahelis et al.
Molecular characterization and molasses fermentation performance of a wild yeast strain operating in an extremely wide temperature range
Bioresour. Technol.
(2009)
Y. Kourkoutas et al.
High temperature alcoholic fermentation of whey using Kluyveromyces marxianus IMB3 yeast immobilized on delignified cellulosic material
Bioresour. Technol.
(2002)
Y. Kourkoutas et al.
Immobilization technologies and support materials suitable in alcohol beverages production: a review
Food Microbiol.
(2004)
A. Mallouchos et al.
Ambient and low temperature winemaking by immobilized cells on brewer’s spent grains: effect on volatile composition
Food Chem.
(2007)
A. Peña-Alvarez et al.
Determination of terpenes in tequila by solid phase microextraction–gas chromatography–mass spectrometry
J. Chrom. A.
(2006)
C. Plata et al.
Formation of ethyl acetate and isoamyl acetate by various species of wine yeasts
Food Microbiol.
(2003)
E. Polychroniadou et al.
Grape and apple wines volatile fermentation products and possible relation to spoilage
Bioresour. Technol.
(2003)

J.A. Regodón Mateos et al.

Influence of Saccharomyces cerevisiae yeast strain on the major volatile compounds of wine

Enzyme Microb. Technol.

(2006)

A. Tsakiris et al.

Immobilization of yeast on dried raisin berries for use in dry white wine-making

Food Chem.

(2004)

R. Vidrih et al.

Synthesis of higher alcohols during cider processing

Food Chem.

(1999)

M. Vilanova et al.

Determination of free and bound terpene compounds in Albariño wine

J. Food Comp. Anal.

(2006)

A. Antonelli et al.

Yeast influence on volatile composition of wines

J. Agric. Food Chem.

(1999)

Cited by (38)

Immobilized cell bioreactors
2022, Current Developments in Biotechnology and Bioengineering: Advances in Bioprocess Engineering
This chapter comprises an integrative contribution to immobilized cell bioreactors technology. A short literature review is given in the introduction, followed by (1) the advances on cell immobilization technology, (2) immobilized cell bioreactors, and (3) their industrial developments. Finally, (4) the chapter makes a critique of immobilized cell bioreactors via a critical review investigation. Specifically, the effects of supports on kinetics and extremely low temperature fermentation, composite supports in relation to two- and three-layer fermentation, and the role of cell immobilization in cell factories development without genetic modifications are discussed. Likewise, the role of tubular cellulose and bacterial cellulose as promoter supports in fermentation are reported. Furthermore, supports and bioreactors, in correlation with multistage fixed bed tower bioreactor development and its applications, are reviewed extensively. Moreover, the cell immobilized bioreactors and their effects on industrial economic analysis, cost, and investment are also discussed. The cell immobilized bioreactors for low- and high-capacity bioprocessing and the industrialization of the technology are important subjects of this chapter. Due to the cell immobilized bioreactor being a technology suitable for beverage production, its effects on wine and beer quality is examined. Finally, an extensive critique of technology is performed of immobilization bioprocessing versus genetic modification, of immobilized cell bioreactor applications, and of the perspectives on nanobiotechnology applications.
Evolution of volatile profile and aroma potential of table grape Hutai-8 during berry ripening
2021, Food Research International
Hutai-8 (Vitis vinifera × Vitis labrusca) is a table grape widely cultivated in China. In order to determine the changes in volatile profile and aroma potential during berry ripening, a total of 84 free and 73 bound aroma compounds were identified and quantified using headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS). Aldehydes and esters were found to be the main volatile compounds in Hutai-8. They accumulated up to 70 days after flowering (DAF) and then decreased. Bound esters and alcohols were prominent. The concentration of bound esters in Hutai-8 at DAF80 was 714.90 µg/L. β-Damascenone, hexanal, (E)-2-hexenal, (E,Z)-2,6-nonadienal, (E)-2-nonenal, and ethyl octanoate significantly contributed to the volatile profile of Hutai-8. The odor activity value (OAV) of hexanal was the highest at DAF80, reaching 351.51. β-Damascenone mainly appeared midway through the maturation process, reaching a concentration of 12.79 µg/L. The majority of free components reached a maximum in DAF70, while the bound components continuously accumulated throughout the mature period. These results suggest that in addition to being a table grape, Hutai-8 has potential for brewing and other processing.
Winemaking using immobilized kefir cells on natural zeolites
2020, LWT
The present study investigated the use of natural zeolites, an inorganic support with multiple technological advantages, for kefir culture immobilization and subsequent evaluation in winemaking at various temperatures (5–45 °C). Cell immobilization was confirmed by Scanning Electron Microscopy and immobilized kefir cells biodiversity was studied by Next Generation Sequencing. Simultaneous alcoholic and malolactic wine fermentations were monitored in repeated batch fermentations for a period >3 months, indicating a high operational stability, even at 5 °C. Alcohol content up to 12.5% vol, malic acid conversion up to 69.2% and ethanol productivity values several fold higher than those observed in traditional practice were reported in fermentations with immobilized cells on natural zeolites. PCA applied to minor volatiles detected by HS-SPME GC/MS showed that mostly the fermentation temperature affected the profile of the aroma-related compounds, while the high total ester concentrations detected at 20 ^οC ascertained the high quality of the products. Importantly, all new wines were accepted by the tasting panel. The results indicated the high potential of immobilized kefir culture on natural zeolites in scale-up industrial applications and suggested their exploitation in the development of low alcohol wines and wines with a semi-sweet character.
Wine production using free and immobilized kefir culture on natural supports
2019, Food Chemistry
The aim of the present study was to perform simultaneous alcoholic and malolactic wine fermentations using free or immobilized kefir culture at a wide temperature range (5–45 °C). Repeated batch fermentations were carried out for a period up to 29 months, suggesting a high operational stability of the systems, while malic acid conversion and ethanol productivity up to 70.9% and 36.9 g/(Ld) were noted. Volatile acidity was at levels typically found in wines (<1.0 g acetic acid/L) in most cases, although increased values were recorded in wines produced at 5 °C, but no vinegar taint was detected. Fusel alcohols were at levels usually found in wines, but were reduced at 5 °C. Application of PCA to minor volatiles showed that the fermentation temperature rather than the nature of kefir culture had a significant effect. Noticeably, all products were accepted by the panel during the preliminary sensory evaluation.
Novel probiotic whey cheese with immobilized lactobacilli on casein
2017, LWT
In the present study, the use of thermally-dried immobilized lactobacilli (Lactobacillus casei ATCC 393 and Lactobacillus delbrueckii ssp. bulgaricus ATCC 11842) on casein was evaluated for the production of a novel dried whey cheese. Cheeses with thermally-dried free lactobacilli cells and with no starter culture were also produced, for comparison reasons. The main physicochemical and microbiological characteristics of a novel whey cheese were determined during ripening at 20 °C until day 60. The results clearly demonstrated that the use of free and immobilized L. casei and L. bulgaricus on casein positively affected the production of dried whey cheeses. More specifically, their use led to novel whey cheeses with improved quality, as resulted by sensory evaluation and SPME GC/MS analysis of aroma related compounds, extended preservation time and protection from spoilage and pathogens. Whey cheeses microbiota was dominated by lactobacilli and mesophilic and thermophilic lactococci. Counts of coliforms, enterobacteria and staphylococci were significantly reduced in whey cheese produced with thermally-dried lactobacilli cultures. The use of cultures, like those proposed in the present study, may have a great potential in dairy technology for the conversion of low quality whey cheeses to novel probiotic foods with improved characteristics.
Assessment of free and immobilized kefir culture in simultaneous alcoholic and malolactic cider fermentations
2017, LWT
The aim of the present study was to assess application of free or immobilized kefir culture on apple pieces and delignified cellulosic material (DCM) in simultaneous alcoholic and malolactic cider fermentations at a wide temperature range (5–45 °C). Repeated batch fermentations were continued for higher than 7 months, showing a high operational stability of the systems and were completed in less than 24 h with immobilized cells on DCM at 37 °C. Malic acid conversion up to 71.5% and ethanol productivity values up to 56.9 g/(Ld) were recorded, which could be adopted by the industrial sector. PCR-DGGE analysis of kefir culture showed no changes in microbial diversity during cell immobilization and fermentations. Analysis of major volatiles documented a significant increase of ethyl acetate content and a decrease of higher alcohols at low temperatures. HS-SPME GC/MS analysis revealed that the highest content of esters and total volatiles was observed in cider fermented by kefir culture immobilized on apple pieces at 20 °C. Principal Component Analysis showed that mainly the immobilization support rather than the temperature had a significant effect on volatile composition. Finally, the sensory evaluation ascertained the high quality of the new products.

View all citing articles on Scopus

View full text

Scale-up of extremely low temperature fermentations of grape must by wheat supported yeast cells

Abstract

Introduction

Section snippets

Yeast strain and media

Results and discussion

Conclusions

Acknowledgement

Bioresour. Technol.

Process Biochem.

Bioresour. Technol.

Bioresour. Technol.

Bioresour. Technol.

Food Microbiol.

Food Chem.

J. Chrom. A.

Food Microbiol.

Bioresour. Technol.

Enzyme Microb. Technol.

Food Chem.

Food Chem.

J. Food Comp. Anal.

Yeast influence on volatile composition of wines

J. Agric. Food Chem.