PRO G max Probiotic fermented soybean meal Benefits of PRO G max

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1 PRO G max Probiotic fermented soybean meal Benefits of PRO G max Probiotic bacteria > CFU/kg High protein with low molecular weight protein approaching small peptides enhancing digestion and absorption of amino acids High protein solubility Reducing anti-nutritional factors in soy bean such as Oligosaccharides, Trypsin inhibitor, Lectin and β-conglycinin

2 PRO G max Probiotic fermented soybean meal PRO G max is produced by solid state fermentation of soy bean meal using high protease bacteria that have the ability to digest large protein molecules in soy bean to small peptides and free amino acids resulted in increasing protein solubility. In addition the nutritional and biochemical characteristics of fermented soy bean meal (FSBM) are also improved by reduction of anti-nutritional factors such as oligosaccharides, trypsin inhibitor, lectin and -conglycinin by action of the bacteria. The protein quality of PRO G max is equivalent to or better than soy protein concentrate (SCP). Probiotic bacteria PRO G max 1 kilogram contains at least or 10 billion cells of living Bacillus subtilis. This bacterium is categorized as one of the probiotics with the excellent properties in tolerant to high temperature and high growth rate at normal conditions as well as in sea water. Bacillus subtilis have been used in preparation of traditional fermented soy bean products all over Asia, for example natto (Japan), thua-nao (Thailand), etc. This Gram positive rod shape bacterium typically produces extracellular proteases, amylases and several other enzymes to digest large biomolecules. The abilities of proteases and amylases can be easily determined by cultivation them on skimmed milk agar and starch agar, respectively.

3 Ability of Bacillus subtilis in producing proteases as evidence by clear zones around the bacterial colonies on skimmed milk agar. Ability of Bacillus subtilis in producing amylases as evidence by clear zones around the bacterial colonies (after iodine staining) on starch agar. Benefits of low molecular weight proteins and small peptides Numerous reports indicated that animal feeds containing low molecular weight proteins fed to young animals were readily absorbed resulted in increasing feed efficiency. Pictures below shows molecular sizes of protein analyzed by SDS-PAGE technique. It is clearly seen that the proteins of PRO G max are relatively small with small peptides comparable to fish meal and skimmed milk which are recognized as high protein quality for animal feed.

4 Solubility of small peptides For animals all nutrients need to be in soluble forms in order to be easily utilized in digestive system. Therefore, high soluble compounds are immediately absorbed in the gastrointestinal tract. The non-soluble substances must be digested to smaller units prior to be available for absorption. The following figure compares the solubility in water of soy bean meal (left-hand tube) and PRO G max (right-hand tube) before milling. It is clearly seen that soy bean meal is insoluble indicated by sedimentation of the meal and clear water while PRO G max is easily soluble in water. Soy bean meal PRO G max Analysis of soluble peptide Proteins are large biological molecules of long chains amino acid residues connecting by peptide bonds. A protein contains at least one long polypeptide. Short polypeptides, containing less than about residues, are commonly called peptides. Bradford method is normally used to determine solubility of proteins by measuring absorbance values of the blue complex compound derived from reaction of proteins and Coomassie dye (Bradford s reagent). Figure shows the low intensity of blue color obtained from soy bean meal extract (left-hand tube) and brilliant blue color obtained from PRO G max (right-hand tube) after reaction with Bradford s reagent. The amounts of soluble peptides were 2.0 g / 100 g and 7.0 g / 100 g, respectively. Soy bean meal PRO G max

5 Reduction of oligosaccharides (Stachyose and Raffinose in soy bean Some organic compounds in soy bean such as non-starch polysaccharides (NSP) cannot be digested and utilized by mono-gastric animals. These NSP compounds are galactooligosaccharides mainly Stachyose and Raffinose. Studies revealed that soy bean meal containing oligosaccharides more that 5% demonstrated harmful effects to young animals because they are unable to digest in small intestine due to lack of endogenous enzymes. In our fermentation process of PRO G max Bacillus subtilis hydrolyzed these oligosaccharides to their basic units, i.e., glucose and galactose as shown in the picture of HPLC chromatograms resulted in effectively reduced Stachyose and Raffinose contents. Soy bean meal PRO G max Nutritional characteristics: Proximate composition and others Protein (min.) 49.0 % ph Moisture (max.) 12.0 % Molecular size of protein Small peptide Fat (max.) 4.0 % Peptide solubility (min.) 7.0 g/100 g Crude Fiber (max.) 4.0 % Pepsin digestibility (min.) 95 % Ash (max.) 7.0 % Bacillus subtilis (min.) 1.0x10 10 CFU/kg

6 Amino acid profile Arginine* 4.01% Aspartic acid 5.44% Histidine* 1.79% Serine 2.83% Isoleucine* 1.88% Proline 2.61% Leucine* 3.54% Glycine 2.10% Lysine* 3.40% Alanine 2.53% Methionine* 0.80% Cystine 0.94% Threonine* 2.20% Tyrosine 1.81% Tryptophan* 0.85% Phenylalanine 2.45% Valine* 2.00% Glutamic acid 9.35% * Essential amino acid Production Process Selected Bacillus subtilis High quality de-hulled Soybean Meal Solid state fermentation Medium temperature drying Fermented soybean meal Grinding Fermented soybean powder: Quality control Packing PRO G max

7 Company Profile Pro Biotech Feed, Co. Ltd. locates at 91/1 Moo 1, Tambon Nakhok, Amphur Mueang, Samut Sakorn 74000, Thailand. The company manufactures and distributes products of protein supplements for animal feed from soy bean meal through fermentation processes by useful microorganisms. Some beneficial bacteria such as Lactobacillus sp. and Bacillus subtilis used in our process known as probiotics are obtained from our commitments through continuous research and development for better and sustainable production of our products. We will continue to improve the quality of our products to serve the animal feed industries which will consequently benefit the farmers. We assure to research and develop new products to serve needs and satisfaction of our valued customers. Pro Biotech Feed, Co. Ltd. production processes conform to the guidelines recommended by Thai government agencies for safe manufacturing of high quality animal feeds and receive GMP (Good Manufacturing Practice) standards as well as HACCP (Hazard Analysis and Critical Control Points) from Department of Livestock Development. Our quality control laboratory staffs strictly monitor the production process to ensure high quality products, free from pathogenic microorganisms and safe for animal consumption. Contact us Pro Biotech Feed, Co. Ltd. 91/1 Moo1, Tambon Nakhok, Amphur Mueang, Samut Sakorn 74000, Thailand. Tel Fax.:

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