Production of buns, the bakery-based snack food, with reduced refined wheat flour content: Recent developments.

Buns are very soft puffed bakery snack items, popular in many countries, especially low- and middle-income nations. Buns are either eaten directly or used in the preparation of culinary items. Buns are mainly prepared using refined wheat flour rich in gluten protein and devoid of husk. Consuming gluten-containing foods is leading to several health complications among consumers worldwide. Hence, several researchers have tried to reduce the gluten content in the dough by incorporating cereals flours, protein-rich sources like soy, cheese whey, etc., hydrocolloids, millets, pomace, and seed flour of vegetables and fruits, etc. These additives not only reduce gluten content in the buns to a certain extent but also enhance the fibre content and nutritional profile of the buns. This mini-review summarizes the recent developments in the production of buns using these additives to improve their nutritional quality.

In-vitro evaluation of antimicrobial and insect repellent potential of supercritical-carbon dioxide (SCF-CO2) extracts of selected botanicals against stored product pests and foodborne pathogens.

In the present study, the antimicrobial and the insect repellent activity of 16 botanical extracts obtained by supercritical CO2 (SCF) extraction were evaluated. The present investigation was conducted as there is a necessity for exploration of natural botanical extracts that target both stored product insects and microbes. The antimicrobial activity was studied by disc diffusion and broth microdilution methods against ten microbial species, including Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis and Listeria monocytogenes), Gram-negative bacteria (Escherichia coli and Salmonella enterica), and fungi (Aspergillus flavus, Aspergillus paraciticus, Aspergillus ochraceous, Aspergillus niger and Penicillium verrucosum). Repellency assay was carried out by area preference method against three coleopteran insects (Tribolium castaneum, Rhyzopertha dominica and Sitophilus oryzae). Among all the extracts, thyme and ajwain were effective against all the tested bacteria with a minimum inhibition concentration (MIC) of 256-1024 µg/mL. Hop extract resulted in better antibacterial activity against all the tested Gram-positive bacteria with a MIC of 32-64 µg/mL. Oregano, thyme and ajwain extracts showed broad-spectrum antifungal activity against all the tested fungi with MIC of 128-1024 µg/mL. Most of the extracts exhibited class V (80.1-100%) repellency against T. castaneum. Extracts of hop, ajwain and thyme were found to have strong repellency against T. castaneum and R. dominica. Therefore, SCF extracts of ajwain and thyme can be explored further for the application of bio-extracts as a growth limiting factors in a microcosm where such consortia thrive.

Development of a carotenoid enriched probiotic yogurt from fresh biomass of Spirulina and its characterization.

Incorporation of Spirulina in milk as thermally dried powder has the disadvantages of non-uniform distribution with undesirable odor and flavor. Through homogenization (200 ± 10 bar), complete dispersion of fresh Spirulina biomass (7% w/w) in milk was achieved and thereafter a carotenoid enriched probiotic yogurt was developed. Confocal microscopy revealed porous Spirulina-milk protein matrix integrated with smaller fat globules in the yogurt. Spirulina led to a 29.56% increase in Lactobacillus acidophilus count, a 20% reduction in fermentation time and a total probiotic count of 1.2 × 107 CFU mL-1. The protein, total chlorophyll, total carotenoid and ß-carotene content (on dry w/w basis) were 3.58 ± 0.08 g 100 g-1, 0.407 ± 0.018 mg g-1, 0.235 ± 0.016 mg g-1 and 13.28 ± 0.08 µg g-1, respectively. During storage (18 days at 6-8 °C), the L. acidophilus count reached 8.83 ± 0.11 log CFU mL-1 with 103.03% increase in the viability by day three and the yogurt retained 71.5% carotenoids. The probiotc Spirulina yogurt was found to be acceptable to consumers as evaluated by affective consumer test.

Fed-Batch Strategies for Production of PHA Using a Native Isolate of Halomonas venusta KT832796 Strain.

In this study, polyhydroxyalkanoates (PHA) accumulation by Halomonas venusta KT832796, a moderate halophilic bacteria isolated from marine source was studied. Both nutritional requirements and process parameters for submerged cultivation of the organism in bioreactor have been standardized. From the shake flask studies, glucose and ammonium citrate as carbon and nitrogen source produced maximum PHA at a ratio 20 with 3.52 g/L of dry cell weight and 70.56% of PHA content. However, ammonium sulfate as the nitrogen source was found to be more suitable for fed-batch cultivation. Several feeding strategies including pH-based fed-batch and variants of pulse feeding were studied to improve the PHA levels. pH-based feeding, although improved PHA level to 26 g/L, most of the carbon flux was diverted towards biomass formation; hence, the percent PHA was only 39.15% of the dry cell weight. Maximum PHA of 33.4 g/L, which corresponded to 88.12% of the dry cell, was obtained from high concentration single pulse method. There was a net 8.65-fold increase in PHA using this feeding strategy when compared to batch studies. According to our knowledge, this is the highest amount of PHA reported for a Halomonas venusta strain.

Betalains rich Rivina humilis L. berry extract as natural colorant in product (fruit spread and RTS beverage) development.

Rivina humilis L. (Phytolaccaceae) or pigeon berry accumulates betalains in its berries. It is reported that the berries are safe to consume, rich in nutrient content and exhibit efficient biological activity. In this report, Rivina berry extract was used as natural colorant in fruit spread and beverage to evaluate its effect on physicochemical properties and acceptability of the product. Results showed that 68 % color retained in Rivina banana spread after 6 months of storage at 5 °C, though there was reduction in L, a and chroma values. Rivina banana beverage lost redness completely during processing. Microbial analysis of the products indicated that they were safe for consumption. The spread had good overall sensorial quality and was liked by consumers indicating that addition of Rivina berry extract did not alter product quality.

Performance evaluation of bulk freeze dried starter cultures of dahi and yoghurt along with probiotic strains in standardized milk of cow and buffalo.

Performance of bulk freeze dried (BFD) cultures of dahi (D) and yoghurt (Y) either with or without probiotic cultures (AB -Lactobacillus acidophilus and Bifidobacterium bifidum) in standardized milk of cow and buffalo was evaluated. In buffalo milk, significantly (p < 0.05) low viable count of probiotic culture combination of dahi cultures (DAB) over non probiotic combination (D) was noticed; whereas, difference in counts of yoghurt culture combinations Y and YAB was not significant. The culture activity of D and DAB was similar in both types of milk, however, the volatile acidity (VA) produced by combination D was higher (32.5 ml/50 g sample) in buffalo milk than in cow milk (29.2 ml/50 g sample). Whereas, DAB produced very low amount of VA (16 ml/50 g sample) both in cow and buffalo milk. The diacetyl and tyrosine contents produced by either D or DAB in cow or buffalo milk were in the same order. Although Y and YAB produced slightly more VA in buffalo milk than in cow milk, significant change in the performance of yoghurt cultures (Y or YAB) both in cow and buffalo milk was not noticed. However, the VA and acetaldehyde produced by YAB either in cow and buffalo milk was higher than that by combination Y. Addition of probiotic cultures significantly enhanced the production of acetaldehyde content in both types of milk. Difference in tyrosine content in yoghurt made either with cow or buffalo milk was not significant. Overall, the present study indicated that the BFD cultures can be used to prepare dahi or yoghurt either from cow or buffalo milk, without affecting the biochemical profile of these products.

Film forming microbial biopolymers for commercial applications--a review.

Microorganisms synthesize intracellular, structural and extracellular polymers also referred to as biopolymers for their function and survival. These biopolymers play specific roles as energy reserve materials, protective agents, aid in cell functioning, the establishment of symbiosis, osmotic adaptation and support the microbial genera to function, adapt, multiply and survive efficiently under changing environmental conditions. Viscosifying, gelling and film forming properties of these have been exploited for specific significant applications in food and allied industries. Intensive research activities and recent achievements in relevant and important research fields of global interest regarding film forming microbial biopolymers is the subject of this review. Microbial polymers such as pullulan, kefiran, bacterial cellulose (BC), gellan and levan are placed under the category of exopolysaccharides (EPS) and have several other functional properties including film formation, which can be used for various applications in food and allied industries. In addition to EPS, innumerable bacterial genera are found to synthesis carbon energy reserves in their cells known as polyhydroxyalkanoates (PHAs), microbial polyesters, which can be extruded into films with excellent moisture and oxygen barrier properties. Blow moldable biopolymers like PHA along with polylactic acid (PLA) synthesized chemically in vitro using lactic acid (LA), which is produced by LA bacteria through fermentation, are projected as biodegradable polymers of the future for packaging applications. Designing and creating of new property based on requirements through controlled synthesis can lead to improvement in properties of existing polysaccharides and create novel biopolymers of great commercial interest and value for wider applications. Incorporation of antimicrobials such as bacteriocins or silver and copper nanoparticles can enhance the functionality of polymer films especially in food packaging applications either in the form of coatings or wrappings. Use of EPS in combinations to obtain desired properties can be evaluated to increase the application range. Controlled release of active compounds, bioactive protection and resistance to water can be investigated while developing new technologies to improve the film properties of active packaging and coatings. An holistic approach may be adopted in developing an economical and biodegradable packaging material with acceptable properties. An interdisciplinary approach with new innovations can lead to the development of new composites of these biopolymers to enhance the application range. This current review focuses on linking and consolidation of recent research activities on the production and applications of film forming microbial polymers like EPS, PHA and PLA for commercial applications.

Effect of carbon and nitrogen sources on simultaneous production of α-amylase and green food packaging polymer by Bacillus sp. CFR 67.

In this paper, effect of different carbon and nitrogen sources, including hydrolysates of rice bran and wheat bran, on simultaneous production of α-amylase (for hydrolysis of starch in food systems) and polyhydroxyalkanoates (PHA, a green biopolymer, which can be used as a packing material for foods) by Bacillus sp. CFR 67 was studied by submerged fermentation. Amongst various carbon sources tested, glucose and sucrose supported production of significantly (P < 0.05) higher amount of α-amylase (66 U/ml) and PHA (444 mg/l), respectively. Of the nitrogen sources tested, ammonium acetate and beef extract led to the production of maximum amount of amylase (36 U/ml) and PHA (592 mg/l), respectively. Supplementation of the production medium with wheat bran hydrolysate (50 ml/l) produced significantly higher amounts of amylase (73 U/ml) and PHA (524 mg/l). Thus this study indicated the potential of agro-residues for the production of value added biomolecules, which can reduce the cost of production of these molecules and enables to reduce the pollution mainly caused by the use of non biodegradable plastics.

Agro-industrial residues and starch for growth and co-production of Polyhydroxyalkanoate copolymer and alfa-amylase by Bacillus. SP CFR67îen

Polyhydroxyalkanoates (PHA) and α-amylase (α-1,4 glucan-4-glucanohydrolase, E.C. 3.2.1.1) were co-produced by Bacillus sp. CFR-67 using unhydrolysed corn starch as a substrate. Bacterial growth and polymer production were enhanced with the supplementation of hydrolysates of wheat bran (WBH) or rice bran (RBH) individually or in combination (5-20 g L-1, based on weight of soluble substrates-SS). In batch cultivation, a mixture of WBH and RBH (1:1, 10 g L-1 of SS) along with ammonium acetate (1.75 g L-1) and corn starch (30 g L-1) produced maximum quantity of biomass (10 g L-1) and PHA (5.9 g L-1). The polymer thus produced was a copolymer of polyhydroxybutyrate-co-hydroxyvalerate of 95:5 to 90:10 mol%. Presence of WBH and corn starch (10-50 g L-1) in the medium enhanced fermentative yield of α-amylase (2-40 U mL-1 min-1). The enzyme was active in a wide range of pH (4-9) and temperature (40-60ºC). This is the first report on simultaneous production of copolymer of bacterial PHA and α-amylase from unhydrolysed corn starch and agro-industrial residues as substrates.

Agro-industrial residues and starch for growth and co-production of polyhydroxyalkanoate copolymer and -amylase by Bacillus sp. CFR-67

Polyhydroxyalkanoates (PHA) and -amylase (-1,4 glucan-4-glucanohydrolase, E.C. 3.2.1.1) were co-produced by Bacillus sp. CFR-67 using unhydrolysed corn starch as a substrate. Bacterial growth and polymer production were enhanced with the supplementation of hydrolysates of wheat bran (WBH) or rice bran (RBH) individually or in combination (5-20 g L-1, based on weight of soluble substrates-SS). In batch cultivation, a mixture of WBH and RBH (1:1, 10 g L-1 of SS) along with ammonium acetate (1.75 g L-1) and corn starch (30 g L-1) produced maximum quantity of biomass (10 g L-1) and PHA (5.9 g L-1). The polymer thus produced was a copolymer of polyhydroxybutyrate-co-hydroxyvalerate of 95:5 to 90:10 mol%. Presence of WBH and corn starch (10-50 g L-1) in the medium enhanced fermentative yield of -amylase (2-40 U mL-1 min-1). The enzyme was active in a wide range of pH (4-9) and temperature (40-60ºC). This is the first report on simultaneous production of copolymer of bacterial PHA and -amylase from unhydrolysed corn starch and agro-industrial residues as substrates.

Agro-industrial residues and starch for growth and co-production of polyhydroxyalkanoate copolymer and α-amylase by Bacillus sp. CFR-67.

Polyhydroxyalkanoates (PHA) and α-amylase (α-1,4 glucan-4-glucanohydrolase, E.C. 3.2.1.1) were co-produced by Bacillus sp. CFR-67 using unhydrolysed corn starch as a substrate. Bacterial growth and polymer production were enhanced with the supplementation of hydrolysates of wheat bran (WBH) or rice bran (RBH) individually or in combination (5-20 g L(-1), based on weight of soluble substrates-SS). In batch cultivation, a mixture of WBH and RBH (1:1, 10 g L(-1) of SS) along with ammonium acetate (1.75 g L(-1)) and corn starch (30 g L(-1)) produced maximum quantity of biomass (10 g L(-1)) and PHA (5.9 g L(-1)). The polymer thus produced was a copolymer of polyhydroxybutyrate-co-hydroxyvalerate of 95:5 to 90:10 mol%. Presence of WBH and corn starch (10-50 g L(-1)) in the medium enhanced fermentative yield of α-amylase (2-40 U mL(-1) min(-1)). The enzyme was active in a wide range of pH (4-9) and temperature (40-60°C). This is the first report on simultaneous production of copolymer of bacterial PHA and α-amylase from unhydrolysed corn starch and agro-industrial residues as substrates.

Characterization of a heat stable anti-listerial bacteriocin produced by vancomycin sensitive Enterococcus faecium isolated from idli batter.

Lactic acid bacteria (LAB) are known to produce various types of bacteriocins, ribosomally synthesized polypeptides, which have antibacterial spectrum against many food borne pathogens. Listeria monocytogenes, a pathogenic bacterium, is of particular concern to the food industry because of its ability to grow even at refrigeration temperatures and its tolerance to preservative agents. Some of the bacteriocins of LAB are known to have anti-listerial property. In the present study, the bacteriocin produced by vancomycin sensitive Enterococcus faecium El and J4 isolated from idli batter samples was characterized. The isolates were found to tolerate high temperatures of 60°C for 15 and 30 min and 70°C for 15 min. The bacteriocin was found to be heat stable and had anti-listerial activity. The bacteriocin did not lost anti-listerial activity when treated at 100°C for 30 min or at 121°C for 15 min. The bacteriocin lost its antimicrobial activity after treating with trypsin, protinase-K, protease and peptidase.

Selection of starter cultures for idli batter fermentation and their effect on quality of idlis.

Idli batter samples were prepared using lactic starter cultures like Pediococcus pentosaceus (Pp), Enterococcus faecium MTCC 5153 (Ef), Ent. faecium (IB2 Ef-IB2), individually, along with the yeast culture, Candida versatilis (Cv). Idli batter prepared using Ef and Ef-IB2 cultures gave better results, when evaluated for the rise in batter volume (80 ml), level of CO2 production (23.8%), titratable acidity 2.4-3.5% (lactic acid) and pH 4.3-4.4. Storage stability of batter made with selected starter cultures was determined by analyzing the idlis prepared using the batter stored for 1 and 5 days for texture, nutrient composition and sensory quality. Slight variations in the results were seen among the idlis of different combination of cultures, whereas these results are better than that of the idlis made using naturally fermented idli batter. Sensory profile of idlis prepared using starter cultures had a higher score (3.9-4.4) compared to the control (3.6) for overall acceptability.

Characterisation of the heat-stable bacteriocin-producing and vancomycin-sensitive Pediococcus pentosaceus CFR B19 isolated from beans.

A bacteriocin-producing lactic culture with antilisterial activity was isolated from beans and identified as Pediococcus pentosaceus CFR B19. It was able to grow and produce bacteriocin at 41 °C but not at 50 °C. This isolate was found to be sensitive to vancomycin and produced heat-stable (at 121 °C for 15 min) bacteriocin. Molecular weight of the purified bacteriocin was found to be ∼4.8 kDa. This isolate can be used as a starter culture or co-culture in fermented milk products and the bacteriocin can be used as a natural preservative in various food products.

Physico-chemical characterization of a new heteropolysaccharide produced by a native isolate of heterofermentative Lactobacillus sp. CFR-2182.

A heterofermentative Lactobacillus sp. CFR-2182 was isolated from dahi samples and it was found to produce 8.0 and 20.5 g/L heteropolysaccharide (HePS) in EPS medium (a simplified synthetic medium) and modified MRS broth, respectively, after 72 h at 30 degrees C. The total carbohydrate, reducing sugar and moisture contents of the purified HePS were 74, 10.6 and 2 g, respectively, per 100 g on dry weight basis. The HePS produced in EPS medium had glucose and mannose in 17:1 ratio. The HePS was non-gelling and non-film forming type. It was completely soluble in water and 1 N sodium hydroxide solution. Gel permeation chromatography and HPLC analysis indicated considerable heterogeneity of the HePS, having three fractions with molecular weights ranging from 3.3 x 10(4) to 1.32 x 10(6) Da. The enzymatic hydrolysis of the HePS with pullulanase and alpha-amylase [with alpha(1-->4) linkage] indicated the presence of alpha(1-->6) and traces of alpha(1-->4) linkages, respectively. NMR analysis of the EPS revealed unique chemical shifts.

Characterization of a new acid stable exo-beta-1,3-glucanase of Rhizoctonia solani and its action on microbial polysaccharides.

A new acid stable exo-beta-1,3-glucanase of Rhizoctonia solani purified from a commercial source 'Kitarase-M', by a combination of ammonium sulfate precipitation, ion-exchange and gel filtration methods, had specific activity of 0.26 U/mg protein, Km and Vmax values of 0.78 mg/ml and 0.27 mM/min/mg protein, respectively. It had molecular weight of 62 kDa with optimum activity at 40 degrees C temperature and pH 5.0, with high stability at pH of 3-7. Unique amino acid sequence was found at N-terminal end. The substrate specificity studies confirmed that it is an exo-beta-1,3-glucanase. It could hydrolyze curdlan powder to release glucose.

Production and characterization of bacterial polyhydroxyalkanoate copolymers and evaluation of their blends by fourier transform infrared spectroscopy and scanning electron microscopy.

Rhizobium meliloti produced a copolymer of short chain length polyhydroxyalkanoate (scl-PHA) on sucrose and rice bran oil as carbon substrates. Recombinant Escherichia coli (JC7623ABC1J4), bearing PHA synthesis genes, was used to synthesize short chain length-co-medium chain length PHA (scl-co-mcl-PHA) on glucose and decanoic acid. Fourier transform infrared spectroscopy (FTIR) spectra of the PHAs indicated strong characteristic bands at 1282, 1723, and 2934 cm(-1) for scl-PHA and at 2933 and 2976 cm(-1) for scl-co-mcl-PHA polymer. Differentiation of polyhydroxybutyrate (PHB) and polyhydroxybutyrate-co-hydroxyvalerate-P(HB-co-HV) copolymer was obseverd using FTIR, with absorption bands at 1723 and 1281 for PHB, and at 1738, 1134, 1215 cm(-1) for HV-copolymer. The copolymers were analyzed by GC and (1)H NMR spectroscopy. Films of polymer blends of PHA produced by R. meliloti and recombinant E. coli were prepared using glycerol, polyethylene glycol, polyvinyl acetate, individually (1:1 ratio), to modify the mechanical properties of the films and these films were evaluated by FTIR and scanning electron microscopy.

Optimization of a new heteropolysaccharide production by a native isolate of Leuconostoc sp. CFR-2181.

AIM: This work is aimed at optimizing the production of a new heteropolysaccharide (HePS) of Leuconostoc sp. CFR-2181 by standardizing the fermentation conditions in a low cost semi-synthetic medium. METHODS AND RESULTS: Both nutritional and cultural parameters, such as carbon source and its concentration, initial pH of the exopolysaccharide (EPS) medium, fermentation temperature and fermentation period were optimized. Fermentation of the EPS medium (pH 6.7), containing sucrose at 5% (w/v) and 5% (v/v) inoculum, at 25 degrees C resulted in maximum production of HePS (18.38 g l(-1)) by the isolate in 4 h of fermentation. CONCLUSIONS: The isolate was found to produce good amount of HePS in just 4 h in a low cost semi-synthetic EPS medium. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on rapid production of HePS by any lactic culture, which can significantly reduce the cost of the EPS.

Optimization of polyhydroxybutyrate production by Bacillus sp. CFR 256 with corn steep liquor as a nitrogen source.

Polyhydroxyalkanotes (PHAs), the eco-friendly biopolymers produced by many bacteria, are gaining importance in curtailing the environmental pollution by replacing the non-biodegradable plastics derived from petroleum. The present study was carried out to economize the polyhydroxybutyrate (PHB) production by optimizing the fermentation medium using corn steep liquor (CSL), a by-product of starch processing industry, as a cheap nitrogen source, by Bacillus sp. CFR 256. Response surface methodology (RSM) was used to optimize the fermentation medium using the variables such as corn steep liquor (5-25 g l(-1)), Na(2)HPO(4) 2H(2)O (2.2-6.2 g l(-1)), KH(2)PO(4) (0.5-2.5 g l(-1)), sucrose (5-55 g l(-1)) and inoculum concentration (1-25 ml l(-1)). Central composite rotatable design (CCRD) experiments were carried out to study the complex interactions of the variables.The optimum conditions for maximum PHB production were (g l(-1)): CSL-25, Na(2)HPO(4) 2H(2)O-2.2, KH(2)PO(4) - 0.5, sucrose - 55 and inoculum - 10 (ml l(-1)). After 72 h of fermentation, the amount of PHA produced was 8.20 g l(-1) (51.20% of dry cell biomass). It is the first report on optimization of fermentation medium using CSL as a nitrogen source, for PHB production by Bacillus sp.

Production of exopolysaccharides by Agrobacterium sp. CFR-24 using coconut water - a byproduct of food industry.

AIMS: The work is intended to explore the suitability of underutilized coconut water (a byproduct of food industry) for the production of exopolysaccharides (EPS) by Agrobacterium sp. CFR 24. METHODS AND RESULTS: Besides checking the suitability of coconut water for the production of water-soluble (WS) and water-insoluble (WIS) EPS, certain fermentation parameters, such as initial pH, incubation period and kinetics of EPS production were investigated. The coconut water medium was found to support the production of both types of EPS. The optimal initial pH and temperature was found to be 6.0 and 30 degrees C, respectively. In shake flask (150 rev min(-1)) studies, high-cell density inoculum resulted in the production of 11.50 g l(-1) of WIS-EPS and 4.01 g l(-1) WS-EPS after 72 and 96 h of fermentation, respectively. CONCLUSIONS: Coconut water was found suitable for the production of microbial EPS by Agrobacterium sp. CFR 24 strain. Under optimum conditions, it produced a good amount of WIS-EPS, which is comparable with that of the sucrose medium (11 g l(-1)). SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on the use of coconut water as a fermentation medium for the production of any microbial EPS. Besides producing value-added products, use of this food industry byproduct, which is often being drained out, can significantly reduce the problem of environmental pollution.