An enzymic technique for the quantitation of galactomannan in guar Seeds.
McCleary, B. V. (1981). Lebensmittel-Wissenschaft & Technologie, 14, 56-59.
An enzymic technique has been developed for the rapid and accurate quantitation of the galactomannan content of guar seeds and milling fractions. The technique involves the measurement of the galactose component of galactomannans using galactose dehydrogenase. The galactomannans are converted to galactose and manno-oligosaccharides using partially purified enzymes from a commercial preparation and from germinated guar seeds. Simple procedures have been devised for the preparation of these enzymes. Application of the technique to a number of guar varieties gave values for the galactomannan content ranging from 22.7 to 30.8% of seed weight.
Measurement of total starch in cereal products by amyloglucosidase-alpha-amylase method: collaborative study.
McCleary, B. V., Gibson, T. S. & Mugford, D. C. (1997). Journal of AOAC International, 80, 571-579.
An American Association of Cereal Chemists/AOAC collaborative study was conducted to evaluate the accuracy and reliability of an enzyme assay kit procedure for measurement of total starch in a range of cereal grains and products. The flour sample is incubated at 95 degrees C with thermostable alpha-amylase to catalyze the hydrolysis of starch to maltodextrins, the pH of the slurry is adjusted, and the slurry is treated with a highly purified amyloglucosidase to quantitatively hydrolyze the dextrins to glucose. Glucose is measured with glucose oxidase-peroxidase reagent. Thirty-two collaborators were sent 16 homogeneous test samples as 8 blind duplicates. These samples included chicken feed pellets, white bread, green peas, high-amylose maize starch, white wheat flour, wheat starch, oat bran, and spaghetti. All samples were analyzed by the standard procedure as detailed above; 4 samples (high-amylose maize starch and wheat starch) were also analyzed by a method that requires the samples to be cooked first in dimethyl sulfoxide (DMSO). Relative standard deviations for repeatability (RSD(r)) ranged from 2.1 to 3.9%, and relative standard deviations for reproducibility (RSD(R)) ranged from 2.9 to 5.7%. The RSD(R) value for high amylose maize starch analyzed by the standard (non-DMSO) procedure was 5.7%; the value was reduced to 2.9% when the DMSO procedure was used, and the determined starch values increased from 86.9 to 97.2%.
Measurement of carbohydrates in grain, feed and food.
McCleary, B. V., Charnock, S. J., Rossiter, P. C., O’Shea, M. F., Power, A. M. & Lloyd, R. M. (2006). Journal of the Science of Food and Agriculture, 86(11), 1648-1661.
Procedures for the measurement of starch, starch damage (gelatinised starch), resistant starch and the amylose/amylopectin content of starch, β-glucan, fructan, glucomannan and galactosyl-sucrose oligosaccharides (raffinose, stachyose and verbascose) in plant material, animal feeds and foods are described. Most of these methods have been successfully subjected to interlaboratory evaluation. All methods are based on the use of enzymes either purified by conventional chromatography or produced using molecular biology techniques. Such methods allow specific, accurate and reliable quantification of a particular component. Problems in calculating the actual weight of galactosyl-sucrose oligosaccharides in test samples are discussed in detail.
Metabolic and genetic perturbations accompany the modification of galactomannan in seeds of Medicago truncatula expressing mannan synthase from guar (Cyamopsis tetragonoloba L.).
Naoumkina, M., Vaghchhipawala, S., Tang, Y., Ben, Y., Powell, R. J. & Dixon, R. A. (2008). Plant Biotechnology Journal, 6(6), 619-631.
Galactomannan gums are widely used in the food and oil industries, and there is considerable interest in applying biotechnological approaches to improve their physical properties. A mannan synthase from guar (Cyamopsis tetragonoloba) was expressed under the control of a bean β-phaseolin promoter in transgenic Medicago truncatula. Although the expression of exogenous mannan synthase caused a slight decrease in galactomannan levels in Medicago, the molecular weight and viscosity of the polymer were significantly increased, although the mannose to galactose ratio and degree of polydispersity remained unchanged. At the same time, expression of about 2.8% of the genes was altered significantly in the seeds of transgenic Medicago lines analysed by Affymetrix genome chip, with a particularly striking induction of putative trehalose phosphate synthase genes. Mannan synthase expression also caused large alterations in the levels of a number of sugars and sugar alcohols, suggesting that over-expression of a processive glycosyltransferase perturbs the mechanisms of sugar sensing and/or homeostasis, possibly involving signalling via trehalose-6-phosphate.
Enzymatic improvement of guar‐based thickener for better‐quality silk screen printing.
Baldaro, E., Gallucci, M., Formantici, C., Issi, L., Cheroni, S. & Galante, Y. M. (2012). Coloration Technology, 128(4), 315-322.
Guar galactomannan (referred to as guar gum) is a versatile polysaccharide, obtained from the seeds of the shrub Cyamopsis tetragonolobus, which finds several applications in either its native or chemically modified form. For textile printing, guar gum can also be partially depolymerised in order to promote dye penetration, improve swelling in water and achieve the desired rheological properties. Guar gum is obtained from guar seeds by a thermo-mechanical process that leaves ca. 3% of largely insoluble proteins in the gum, originating from the endosperms aleurone layer. When printing silk fabrics with acid or premetallised dyes, guar endogenous insoluble proteins bind tightly to anionic dyes, causing deposition of coloured aggregates on the fabric. This causes imperfections on the printed fabric in the form of tiny, but visible, ‘dots’, which lowers the quality of the final articles. In order to eliminate ‘dotting’, a novel printing thickener composed of depolymerised guar gum mixed with a bioengineered subtilisin protease has been developed. Upon solubilisation of the gum, and during preparation of the printing paste mixture, the protease hydrolyses guar gum insoluble proteins, generating soluble peptides that are washed off by the post-printing treatments of the fabric. This enzymatic application prevents ‘dotting’ and significantly improves the quality of the silk print, without any measurable tensile strength loss of the fabric.
Nutrient utilisation and intestinal fermentation are differentially affected by the consumption of resistant starch varieties and conventional fibres in pigs.
Rideout, T. C., Liu, Q., Wood, P. & Fan, M. Z. (2008). British Journal of Nutrition, 99(05), 984-992.
This study examined the influence of different resistant starch (RS) varieties and conventional fibres on the efficiency of nutrient utilisation and intestinal fermentation in pigs. Thirty-six pigs (30 kg) were fed poultry meal-based diets supplemented with 10 % granular resistant corn starch (GCS), granular resistant potato starch (GPS), retrograded resistant corn starch (RCS), guar gum (GG) or cellulose for 36 d according to a completely randomised block design. Distal ileal and total tract recoveries were similar (P> 0•05) among the RS varieties. Distal ileal starch recovery was higher (P< 0•05) in pigs consuming the RS diets (27–42 %) as compared with the control group (0•64 %). Consumption of GCS reduced (P< 0•05) apparent total tract digestibility and whole-body retention of crude protein in comparison with the control group. Consumption of GPS reduced (P< 0•05) total tract Ca digestibility and whole-body retention of Ca and P compared with the control group. However, consumption of RCS increased (P< 0•05) total tract Ca digestibility compared with the control group. Caecal butyrate concentration was increased (P< 0•05) following consumption of RCS and GG in comparison with the control group. Consumption of all the RS varieties reduced (P< 0•05) caecal indole concentrations compared with the control. Caecal butyrate concentrations were positively correlated (P< 0•05; r 0•63–0•83) with thermal properties among the RS varieties. We conclude that nutrient utilisation and intestinal fermentation are differentially affected by the consumption of different RS varieties and types of fibres. Thermal properties associated with different RS varieties may be useful markers for developing RS varieties with specific functionality.
Seed yield, galactomannan content and quality traits of different guar (Cyamopsis tetragonoloba L.) genotypes.
Gresta, F., Ceravolo, G., Presti, V. L., D’Agata, A., Rao, R. & Chiofalo, B. (2017). Industrial Crops and Products, 107, 122-129.
The aim of this study was to assess the productive and qualitative traits of seeds of six guar genotypes coming from India, South Africa and the USA, grown in a Mediterranean environment, both for industrial use (galactomannans) and as animal feed (guar meal). Yield of the six varieties ranged from 1.49 t ha-1 to 2.05 t ha-1. Galactomannan content reached values from 28.6 g/100 g to 34.6 g/100 g with the highest significative value exhibited by Indian genotype, Lewis and South African genotype. Galactomannan yield resulted 0.55 t ha-1, on average, with small differences among genotypes. The mean values of protein content of the guar seeds showed an average value of all the genotypes of 283 g/kg on a DM basis of 896 g/kg with the significantly highest values in Indian and South African genotypes. Lipids (on a DM basis) was of 28.3 g/kg DM on average, with the significantly highest value in South African and Indian genotypes. Crude Fibre, ranged from 107 to 122 g/kg. The fibrous fractions were on average 496 g/kg DM, 133 g/kg DM and 5.08 g/kg DM for NDF, ADF and ADL, respectively. Polyphenols ranged from 3.61 mg gallic acid/g of Matador to 6.63 mg gallic acid/g of Kinman and tannins from 2.01 mg/g of catechin equivalents of Kinman to 4.05 mg/g of catechin equivalents of South African genotype. Monument showed the highest content of n6-PUFA, whereas, the highest content of n3-PUFA was observed in Indian genotype. The ratio n3/n6 PUFA, the Atherogenic and Thrombogenic indices underline the high quality of guar oil for human and animal nutrition. This study underlines the double aptitude of guar seed of gum extraction and the by-product, assessing, respectively, its productive traits for industrial use and nutritional traits for increasing its use in feedstuff in the Mediterranean crop-livestock system. This double use represents an important added value to promote the widespread cultivation of the crop.
Galactomannan from Trigonella foenum‐graecum L. seed: Prebiotic application and its fermentation by the probiotic Bacillus coagulans strain MTCC 5856.
Majeed, M., Majeed, S., Nagabhushanam, K., Arumugam, S., Natarajan, S., Beede, K. & Ali, F. (2018). Food Science & Nutrition, In Press.
Health benefits of dietary fibers are currently being widely recognized. However, the assessment of dietary fiber as a prebiotic is essential and also important for the development of an improved synbiotic commercial preparation. Thus, the aim of this study was to evaluate the potential of galactomannan extracted from fenugreek seeds as a prebiotic fiber and also its fermentation by the probiotic strain Bacillus coagulansMTCC 5856. Nondigestibility by the gastric acid and pancreatic enzyme hydrolysis of galactomannan were determined using an in vitro model mimicking the in vivo conditions. Further, anaerobic fermentation and utilization of galactomannan by the B. coagulansMTCC 5856 was investigated followed by selective inhibition of Escherichia coliATCC 25922. The galactomannan from fenugreek seeds was found to be nondigestible to gastric acid and also to pancreatic enzymatic hydrolysis. The galactomannan was fermented and utilized (71.4%) by the B. coagulansMTCC 5856, and also significant amount of short-chain fatty acids production was also observed. Furthermore, B. coagulansMTCC 5856 inhibited the E. coliATCC 25922 growth when cocultured with galactomannan suggesting competitive fermentation of probiotic bacteria. Galactomannan exhibited prebiotic activity and also showed suitability with probiotic B. coagulansMTCC 5856 in a synbiotic combination. This study provides the first scientific evidence of galactomannan from fenugreek seeds as a prebiotic that may play an important role in modulating gut flora by acting as substrate to beneficial microbes.
Lipopolysaccharide immune stimulation but not β-mannanase supplementation affects maintenance energy requirements in young weaned pigs.
Huntley, N. F., Nyachoti, C. M. & Patience, J. F. (2018). Journal of Animal Science and Biotechnology, 9(1), 47.
Background: Pathogen or diet-induced immune activation can partition energy and nutrients away from growth, but clear relationships between immune responses and the direction and magnitude of energy partitioning responses have yet to be elucidated. The objectives were to determine how β-mannanase supplementation and lipopolysaccharide (LPS) immune stimulation affect maintenance energy requirements (MEm) and to characterize immune parameters, digestibility, growth performance, and energy balance. Methods: In a randomized complete block design, 30 young weaned pigs were assigned to either the control treatment (CON; basal corn, soybean meal and soybean hulls diet), the enzyme treatment (ENZ; basal diet + 0.056% β-mannanase), or the immune system stimulation treatment (ISS; basal diet + 0.056% β-mannanase, challenged with repeated increasing doses of Escherichia coli LPS). The experiment consisted of a 10-d adaptation period, 5-d digestibility and nitrogen balance measurement, 22 h of heat production (HP) measurements, and 12 h of fasting HP measurements in indirect calorimetry chambers. The immune challenge consisted of 4 injections of either LPS (ISS) or sterile saline (CON and ENZ), one every 48 h beginning on d 10. Blood was collected pre- and post-challenge for complete blood counts with differential, haptoglobin and mannan binding lectin, 12 cytokines, and glucose and insulin concentrations. Results: Beta-mannanase supplementation did not affect immune status, nutrient digestibility, growth performance, energy balance, or MEm. The ISS treatment induced fever, elevated proinflammatory cytokines and decreased leukocyte concentrations (P < 0.05). The ISS treatment did not impact nitrogen balance or nutrient digestibility (P > 0.10), but increased total HP (21%) and MEm (23%), resulting in decreased lipid deposition (-30%) and average daily gain (-18%) (P < 0.05). Conclusions: This experiment provides novel data on β-mannanase supplementation effects on immune parameters and energy balance in pigs and is the first to directly relate decreased ADG to increased MEm independent of changes in feed intake in immune challenged pigs. Immune stimulation increased energy partitioning to the immune system by 23% which limited lipid deposition and weight gain. Understanding energy and nutrient partitioning in immune-stressed pigs may provide insight into more effective feeding and management strategies.