Title of Invention

A PROCESS FOR THE PREPARATION OF PREBIOTIC BREAD SPREAD

Abstract

The present invention relates to a process for the preparation of prebiotic bread spread. The process uses Fructo-oligosaccharides (FOS) produced using extra cellular fructosyl transferase from Aspergillus oryzae. The process involves optimization of parameters to obtain desirable product. The product contains 50% FOS with functional properties including prebiotic attributes.The novelty of the present invention lies in the use of extra cellular fructosyl tranferase from Aspergillus oryzae CFR 202 for fermentation of bread spread. The novelty of the bread spread is that it is prepared by fermentation process. The product contains 50% FOS with functional properties like non-carcinogenicity, low calorific value and prebiotic property. FOS also improves mineral absorption, reduces that total cholesterol and triglyceride levels in the body. The product does not involve the addition of any chemicals directly. The product can also be enriched with minerals, nutrients, fruits chunks and nuts to improve the organoleptic properties of the product.

Full Text

The present invention relates to a process for the preparation of prebiotic bread spread. The process uses FOS produced using extracellular fructosyl transferase (FTase) from Aspergillus oryzae CFR 202. Bread spreads are one of the most versatile products in the food industry. Spreads and jams based on fruits and fruit products are common in the market and are of great demand. They can be used for various applications and are produced by many small and large scale manufacturers. Even though there are a number of patents relating to the preparation of spreads using various fruits, there are no reports regarding the preparation of a bread spread by fermentation. Reference may be made to a process for the preparation of a reduced calorie fruit spread (Sharp SE and Antenucci RN, US 5270071, December 1993) wherein a reduced calorie fruit spread was prepared by mixing fruit or fruit flavouring with sucralose, methoxy pectin or carrageenan, carboxy methyl cellulose, guar gum and locus bean gum. The process is different from the present invention since it involves the use of fruit or fruit flavoring and many additional ingredients like sweetener, gum etc. In addition, it lacks the prebiotic properties. Reference may be made to a process for the preparation of a honey spread with dried fruits, nuts, spices, natural flavors and colors (Preston M B, and Smith D A, www.nhb.org. Honey spreads, National Honey board, Longmont, Colorado, NHB, 1995) wherein crystallized honey was mixed with dried fruits, nuts, spices and natural flavours and colors. In another embodiment of the process honey fruit spreads were prepared using honey as the major sweetener and natural pectins, gums / stabilizers to produce a stable gel of desirable consistency. This process is different with respect to the starting material and the additives used. It also has high calorific value (3 kCal/g) compared to the product of the present process (1.5 kCal/g) and lacks prebitoic properties. Reference may be made to a process for the preparation of a low fat spread (Schotel, Ronald Albert, US 5624702, October, 1995) wherein a reduced fat spread is prepared having more than 15 wt % inulin, particularly those derived from Jerusalem artichoke or chicory, 0.1 to 15 wt % of mono- or disaccharides and 0.1 to 7 wt % of fat. The process is different since it uses plant-derived polyfructoses and contains additional fat and monosaccharides. Reference may be made to a process for the preparation of a low fat spread comprising of more than 15 wt. % of inulin (an oligofructose) derived from Jerusalem artichoke or chicory and 0.1 to 7 wt. % of fat derived from the group consisting of dairy cream, butter fat and cream cheese (Schotel, Ronald Albert, US 6,348,228, November 1997). The process is different from the present invention since it involves the use of inulin derived from a plant source and not a fermentation product. Another drawback is that the concentration of inulin is low in plants and the mass production is dependent on seasonal conditions. Reference may be made to a process wherein a butter flavoured low-fat spread has been formulated using vegetable fat, soy concentrate and skim milk powder (Patel A A and Gupta S K, Indian Food Industry, Jan- Feb 1993, 12 28- 31). The drawbacks of the product are its high calories (3.92 kCal/g), high fat content and lack of prebiotic properties. The main object of the present invention is to provide a process for the preparation of prebiotic bread spread which obviates the drawbacks outlined above. Another object of the present invention to optimize conditions to obtain the desired bread spread. The deposition number of the organism Aspergillus oryzae is MMTC 5154 deposited at IMTECH,Chandigarh,India. The source of this organism is Soil isolate from the lab and origin is fungal isolate. Accordingly, the present invention provides a process for the preparation of prebiotic bread spread, which comprises; a) growing the culture Aspergillus oryzae in a medium containing 1 % sucrose and 0.2% yeast extract at pH ranging between 5 and 6, at temperature ranging between 25 and 35ºC, b) shaking the above said medium at a speed of 200-250 rpm for a period of 24-48 hr to develop the inoculums, c) transferring 10-25 % of the inoculums to 5-20 g sterilized solid substrate selected from known cereal bran, preferably wheat bran having a moisture content of 40 to 60%, d) incubating the above said mixture for a period of 48-120 hr at a temperature of 25-35ºC, e) suspending the above said moldy substrate in water, f) incubating the flasks containing the suspended moldy substrate on a rotary shaker at a speed of 200-220 rpm for a period of 1-2 hr, g) separating the substrate by filtration to obtain the extract. h) incubating the extract with 60% sugar solution in 0.1 M citrate buffer of pH ranging between 5.0 and 5.5 for a period in the range of 18-20 hr at a temperature in the range of 50 to 60°C and, i) concentrating by heating the above said reaction mixture at a temperature ranging between 75 and 85°C for 2-3 hr under stirring to obtain the concentrated product, followed by cooling for a period of 15-30 min to obtain the desired bread spread. In an embodiment of the present invention, the pH of the buffer used in step lh may be preferably 5.15. The novelty of the present invention lies in the use of extra cellular fructosyl tranferase from Aspergillus oryzae CFR 202 for fermentation of bread spread. The novelty of the bread spread is that it is prepared by fermentation process. The product contains 50% FOS with functional properties like non-carcinogenicity, low calorific value and prebiotic property. FOS also improves mineral absorption, reduces the total cholesterol and triglyceride levels in the body. The product does not involve the addition of any chemicals directly. The product can also be enriched with minerals, nutrients, fruit chunks and nuts to improve the organoleptic properties of the product. The process for the preparation of the bread spread is illustrated in the following flow chart. Five -Eight days old slant of Aspergillus oryzae CFR 202 One loop full of spores transferred to medium containing 1 % sucrose and 0.2 % yeast extract at pH ranging from 5-6 and incubated for 24-48 h at temperature ranging from 25-35 °C at about 200-250 rpm to develop inoculum 10-25 % inoculum transferred I 5-20 g substrate moistened with 2.5-10 ml water Incubated for 48-120 h at temperature ranging from 25-35 °C Moldy substrate extracted with 50-100 ml water after shaking for 1 -2 h at 200 I Extract filtered - 220 rpm Crude Fructosyl Transferase (FTase) FTase (0.25 ml) mixed with 600 g/l sucrose (1 .75 ml) at pH 5-5.5 for 18-20 t h at 50-60°C Reaction mixture concentrated by heating (80 ± 2 °C) for 4-5 h with stirring Cooling the concentrated product for 15 - 30 minutes for setting Packing the spread in sterilized PET bottles or any suitable container The following examples are given by way of illustration of the present invention and therefore should not be construed to limit the scope of the present invention. EXAMPLE - 1 Aspergillus oryzae CFR 202 was grown in 50 ml_ medium consisting of 1 % sucrose and 0.2 % yeast extract (pH 5.5) at 30 °C for 24 h at 250 rpm to develop inoculum. 20 % v/v of the inoculum was transferred to a 250 ml conical flask containing 10 g wheat bran moistened with 10 ml water and incubated for 48 h at 30 °C. The moldy substrate was mixed with 50 ml water and subjected to shaking at 220 rpm for one and a half hours. The extract is then filtered and used as the source of extracellular enzyme for the production of FOS. 240 mL of the extract was mixed with 1 kg of sugar dissolved in 667 ml 0.1 M citrate buffer (pH 5.15) and incubated for 18 h at 55 °C with stirring. The final volume of reaction mixture was 1.5 L (50° brix, pH - 5.32). The reaction mixture was heated at 80 ± 2 °C for 2 h with stirring. The concentrated product (78 ° brix) was analyzed by HPLC using refractive index detector. The concentration of FOS obtained was 50 % of the initial sucrose. The product obtained was poured into a clean sterilized container. EXAMPLE - 2 Aspergillus oryzae CFR 202 was grown in 50 ml medium consisting of 1 % sucrose and 0.2 % yeast extract (pH 5.5) at 30 °C for 24 h at 250 rpm to develop inoculum. 20 % v/v of the inoculum was transferred to a 250 ml conical flask containing 10 g wheat bran moistened with 10 mL water and incubated for 48 h at 30 °C. The moldy substrate was mixed with 50 ml water and subjected to shaking at 220 rpm for one and a half hours. The extract is then filtered and used as the source of extracellular enzyme for the production of FOS. 1.2 L of the extract was mixed with 5 kg of sugar dissolved in 3.35 L 0.1 M citrate buffer (pH 5.15) and the reaction was carried out in a 10 L reactor for 18 h at 55 °C with constant agitation (100 rpm). The final volume of reaction mixture was 8 L (50° brix, pH - 5.32). The reaction mixture was then heated at 80 ± 2 °C for 2 h with stirring. The concentrated product (79 ° brix) had a concentration of FOS corresponding to 50 % of the initial sucrose. The product obtained was poured into a clean sterilized container. The main advantages of the present invention are: 1. The bread spread is prepared by a fermentation process. 2. The product is prebiotic based and contains 50% FOS with functional properties. 3. The product does not involve the use of any chemicals. 4. The product has the potential for supplementation with minerals, nutrients, fruit chunks and nuts to improve the organoleptic properties of the product. We Claim: 1. A process for the preparation of prebiotic bread spread, which comprises: a) growing the culture Aspergillus oryzae in a medium containing 1 % sucrose and 0.2% yeast extract at pH ranging between 5 and 6, at temperature ranging between 25 and 35ºC, b) shaking the above said medium at a speed of 200-250 rpm for a period of 24-48 hr to develop the inoculums, c) transferring 10-25 % of the inoculums to 5-20 g sterilized solid substrate selected from known cereal bran, preferably wheat bran having a moisture content of 40 to 60%, d) incubating the above said mixture for a period of 48-120 hr at a temperature of 25-35°C, e) suspending the above said moldy substrate in water, f) incubating the flasks containing the suspended moldy substrate on a rotary shaker at a speed of 200-220 rpm for a period of 1 -2 hr, g) separating the substrate by filtration to obtain the extract, h) incubating the extract with 60%) sugar solution in 0.1 M citrate buffer of pH ranging between 5.0 and 5.5 for a period in the range of 18-20 hr at a temperature in the range of 50 to 60ºC and, i) concentrating by heating the above said reaction mixture at a temperature ranging between 75 and 85ºC for 2-3 hr under stirring to obtain the concentrated product, followed by cooling for a period of 15-30 min to obtain the desired bread spread. 2. A process as claimed in claim 1, wherein the pH of the buffer used in step h is preferably 5.15. 3. A process for the preparation of periodic bread spread substantially as herein described with reference to the examples.

Documents:

344-del-2002-abstract-(22-10-2008).pdf

344-del-2002-abstract.pdf

344-del-2002-claims-(22-10-2008).pdf

344-del-2002-claims.pdf

344-del-2002-complete specification (granted).pdf

344-DEL-2002-Correspondence-Others-(15-10-2008).pdf

344-del-2002-correspondence-others-(22-10-2008).pdf

344-del-2002-correspondence-others.pdf

344-del-2002-correspondence-po.pdf

344-del-2002-description (complete)-(22-10-2008).pdf

344-del-2002-description (complete).pdf

344-del-2002-form-1.pdf

344-del-2002-form-18.pdf

344-del-2002-form-2.pdf

344-del-2002-form-3.pdf