Lactobacillus casei cell immobilization on fruit pieces for probiotic additive, fermented milk and lactic acid production

doi:10.1016/j.procbio.2004.01.029

Process Biochemistry

Volume 40, Issue 1, January 2005, Pages 411-416

https://doi.org/10.1016/j.procbio.2004.01.029 Get rights and content

Abstract

Lactobacillus casei cells were immobilized on fruit (apple and quince) pieces separately and the immobilized biocatalysts were used for 15 successive fermentation batches of whey immediately after their preparation and for fermentations of milk immediately after their preparation and after a long storage period at low temperature. The immobilized biocatalysts used for successive fermentation batches of whey proved to be very effective and suitable for food grade lactic acid production, while no loss of activity at all temperatures (30, 37, 45 °C) tested was observed, as very high lactose conversion was reported. Cell immobilization of L. casei on apple and quince pieces was also shown by electron microscopy. In addition, apple and quince supported biocatalysts were used for milk fermentation immediately after their preparation, and after storage for 15, 98, and 129 days at 4 °C. No infection was reported during the storage periods. After storage, the immobilized biocatalysts were reactivated very quickly and produced milk fermentation products with a fruity, distinctive aroma which remained during all storage period.

Introduction

Nowadays, a new trend is the development of novel fermented milks, which contain microorganisms, called probiotics such as Lactobacillus acidophilus, Lactobacillus casei, and others. Probiotic lactic acid bacteria act beneficially in human health, and therefore, a wide variety of lactic acid bacteria strains are available to consumers in both traditional fermented foods and in supplement form [1], [2]. Generally, the production of probiotic foods that will contain specific probiotic strains at suitable levels of viable cells during their shelf life is a technological challenge [3].

The shelf life of probiotics should be controlled in order to manufacture products with adequate live bacteria (at least 10⁷ CFU/g) to obtain the health promoting benefits of probiotic cultures [4].

The viability of probiotic bacteria can be improved by methods such as immobilization, appropriate selection of acid, and bile resistant strains, use of oxygen impermeable containers, stress adaptation, and others [2].

Lactic acid is also an important chemical used in a wide variety of applications, being used primarily in the food industry as an acidulant, preservative, and for the production of emulsifying agents [5].

Lactobacillus casei cells have been immobilized in some supports for lactic acid production. Agar was more effective than polyacrylamide for L. casei entrapment for lactic acid production from whey [6]. Also, calcium pectate gel and chemically modified chitosan beads were used as supports for L. casei cell immobilization [7]. Alginate has so far been a popular matrix for immobilization of lactic acid bacteria [8], [9], [10], [11]. Other supports used for immobilization include porous foam glass particles [12], ceramic beads or porous glass [13], poraver beads [14], and gluten pellets [15].

However, cell immobilization on a food-grade support is essential for food production. In addition, aroma and taste play a significant role in customer acceptance.

Apple [16], [17] and quince pieces [18], [19] have been proposed as immobilization supports of yeast strains for room- and low-temperature wine-making. Apple and quince pieces are of food-grade purity, cheap, and abundant in nature, acid resistant supports, while the immobilization technique is simple and easy and showed high operational stability and a significant increase in productivity in alcohol production. The products (wine) were of very good quality with a distinctive aromatic potential. In addition, apple pieces proved to be very effective supports for the survival of apple-immobilized yeast cells, as the immobilized biocatalyst was able to reactivate after storage of 120 days [20].

In order to increase probiotic viability, production of apple and quince pieces supported L. casei is necessary due to the suitability of the supports as food ingredients. The produced biocatalysts have to be also examined for their suitability for lactic acid production. Therefore, the aims of this investigation were the increase of probiotic viability through fruit supported probiotic organism and use of the produced biocatalysts for lactic acid production.

Section snippets

Materials and methods

Lactobacillus casei isolated from a commercial dairy product was used in the present study. It was grown on synthetic medium containing (%, w/v): yeast extract 0.5%, K₂HPO₄ 0.1%, (NH₄)₂SO₄ 0.1%, MgSO₄·7H₂O 0.5%, and lactose 2% in distilled water. This medium was sterilized at 121 °C for 15 min. Flasks were incubated at 37 °C without agitation.

Whey was produced in the laboratory after milk coagulation using the enzyme rennet. It had a pH 6.4–6.6 and contained 50 g lactose/l.

Results and discussion

The experimental part of this investigation was organized in order to show (i) cell immobilization of L. casei on fruit pieces; (ii) survival of fruit pieces supported L. casei; and (iii) efficiency of the produced biocatalysts in probiotic fermented milk and lactic acid production.

Lactobacillus casei cells were immobilized on apple and quince pieces separately and then 15 successive whey fermentation batches for each immobilized biocatalyst were carried out at 30, 37, and 45 °C.

In order to

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Manufacture of fermented lactic beverages containing probiotic cultures

J. Food Sci

(2002)

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Natural or synthesized materials can be used for cell immobilization to enhance cell viability and productivity in fermentation processes (batch, fed-batch or continuous fermentations). 3D printing is one of the newest technologies to produce unique immobilization support materials. In this research, the selection strategy of 3D lattice structure type for unique immobilization materials, effect of different lattice structure types on cell immobilization in a shake flask experiments and optimization of ethanol production in a laboratory scale bioreactor with 3D spheres were studied. Firstly, 3D designs were compared to determine the best type of 3D spheres with a higher area:volume ratio for cell immobilization. Then shake flask experiments were performed and showed that W, 20 mm is the best 3D sphere combination for ethanol fermentation. Finally, over 37 g/L ethanol (ethanol yield ranged from 41.91% to 46.36%) was produced at all repeated batch cycles by using 100 g/L initial sugar with at least 82% sugar utilization capacity in a lab-scale bioreactor successfully. All results showed that 3D-printed materials could be used as immobilization agents for microorganisms by selecting the proper geometry, size and printing technology.
Potential utilization of dairy industries by-products and wastes through microbial processes: A critical review
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Citation Excerpt :
On the contrary, high productivity occurs even in low-volume fermenters when continuous production systems were applied (Aeschlimann and von Stockar, 1991; Panesar et al., 2007; Soriano-Perez et al., 2012). In continuous batch systems, immobilized cells can be successfully applied for lactic acid production using by dairy wastes (Kourkoutas et al., 2005; Panesar and Kennedy, 2012). Comprehensively, lactic acid production from dairy effluents by microbial fermentation has been increased recently.
The dairy industry generates excessive amounts of waste and by-products while it gives a wide range of dairy products. Alternative biotechnological uses of these wastes need to be determined to aerobic and anaerobic treatment systems due to their high chemical oxygen demand (COD) levels and rich nutrient (lactose, protein and fat) contents. This work presents a critical review on the fermentation-engineering aspects based on defining the effective use of dairy effluents in the production of various microbial products such as biofuel, enzyme, organic acid, polymer, biomass production, etc. In addition to microbial processes, techno-economic analyses to the integration of some microbial products into the biorefinery and feasibility of the related processes have been presented. Overall, the inclusion of dairy wastes into the designed microbial processes seems also promising for commercial approaches. Especially the digestion of dairy wastes with cow manure and/or different substrates will provide a positive net present value (NPV) and a payback period (PBP) less than 10 years to the plant in terms of biogas production.
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Lactic acid bacteria (LAB) are a diverse group of microorganisms present in many ecological systems and are a very important part of human nutrition, health, and food preservation. Fermented foods are generally rich in health-promoting microorganisms, which benefit and enhance the well-being. Fermentation has three main purposes: preservation of foods and beverages, the production of nutritionally relevant components, and the improvement of their palatability. LAB are susceptible to numerous stress factors during fermentation, which greatly hinder the production efficiency of the process. Many biotechnological processes greatly benefit from immobilization techniques. The cell encapsulation could be considered as an alternative to conventional processes in order to improve the survival of microorganisms added to different food matrices. Moreover, encapsulates facilitate formulations of food products that are healthier, tastier, and more convenient, and the development of delivery systems for biologically active compounds in food systems is an important issue in modern food technology. Encapsulation of active substances into carrier materials can be attained by several methods. The carrier/wall material should be GRAS, biodegradable, inexpensive, and have the capability of creating a barrier between the core material and the external environment, thus protecting the encapsulated microbial cells in environments that could damage cell.
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Food fermentation has been practiced since prehistoric times and developed over the years with modification in substrates, processes and technologies. It is carried out by microbial cultures with techniques such as back slopping and enrichment, enhancing the organoleptic property, nutrient availability and storage life of food and in many cases contributing beneficial microbes to the diet of the consumer.
The microorganisms involved in fermentation greatly vary according to the food product. One of the major groups of microbes used in traditional and industrial fermentation of cereals includes the lactic acid bacteria, many of which are known to have probiotic characteristics followed by yeast and mould. While dairy fermented foods have received attention as a source of probiotic microbes, those in non-dairy foods especially millets and cereal mixtures have not been documented. Traditional fermented foods of animal origin as well as other plant foods such as pulse, fruits and vegetables are also discussed. The review examines the presumptive probiotic bacteria found in non-dairy fermented foods around the globe.
This review is an attempt to collate the scientific progress with respect to probiotics in millet fermented foods, particularly of Africa and Asia. India has the largest documented number of fermented foods followed by Korea especially made with cereals. These fermented foods containing presumptive probiotic microorganisms may serve as probiotic supplements that are affordable as opposed to expensive probiotic formulations.
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Lactobacillus casei CSL3: Evaluation of supports for cell immobilization, viability during storage in Petit Suisse cheese and passage through gastrointestinal transit in vitro
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The present study predicted an evaluation of supports for immobilization of Lactobacillus casei CSL3, as well as its viability during storage in Petit Suisse cheese and passage through the GIT in vitro.
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Process Biochemistry

Abstract

Introduction

Section snippets

Materials and methods

Results and discussion

References (22)

Probiotic bacteria: selective enumeration and survival in dairy foods

J. Dairy Sci.

Technological challenges for future probiotic foods

Int. Dairy J.

Lactic acid production from whey permeate by immobilized Lactobacillus casei

Enzyme Microb Technol

Malolactic fermentation in Chardonnay wine by immobilized Lactobacillus casei cells

Process Biochem

Encapsulation of Lactobacillus casei cells in liquid-core alginate capsules for lactic acid production

Enzyme Microb Technol

Lactic acid production from deproteinized whey by mixed cultures of free and coimmobilized Lactobacillus casei and Lactococcus lactis cells using fedbatch culture

Enzyme Microb Technol

Lactic acid production by immobilized Lactobacillus casei in recycle batch reactor: a step towards optimization

J Biotechnol

Continuous wine fermentation using a psychrophilic yeast immobilized on apple cuts at different temperatures

Food Microbiol

Wine production using yeast immobilized on quince at temperatures between 30 and 0 °C

Food Chem

Manufacture of fermented lactic beverages containing probiotic cultures

J. Food Sci

Cited by (123)

Ethanol production by immobilized Saccharomyces cerevisiae cells on 3D spheres designed by different lattice structure types

Potential utilization of dairy industries by-products and wastes through microbial processes: A critical review

Encapsulation technology of lactic acid bacteria in food fermentation

Probiotic microorganisms from non-dairy traditional fermented foods

Antioxidant activity of Bifidobacterium animalis MSMC83 and its application in set-style probiotic yoghurt

Lactobacillus casei CSL3: Evaluation of supports for cell immobilization, viability during storage in Petit Suisse cheese and passage through gastrointestinal transit in vitro