Affinity purification of 4-α-glucanotransferase through formation of complex with insoluble amylose

4-α-Glucanotransferases (αGTases) are useful enzymes to modify starch structure using their unique hydrolysis pattern and transglycosylation activity. In spite of lacking of a carbohydrate binding module, a thermostable aGTase from Thermus thermophilus (TTαGT) bound to insoluble amylose, leading to simple recovery of the enzyme-amylose complex using centrifugation. At a preparative scale, the recovery yield was 40.3% of the subjected free enzyme via collection of the complex twice. The complex exhibited improved thermostability, which shifted the optimum temperature from 70 to 80°C and increased the half-life at 90°C by three-fold compared with the free enzyme. Texture profile analysis of the gels made of modified starch by either free TTαGT or the complex revealed that the complex with double dosage showed the performance similar to free TTαGT in the modification of starch. In conclusion, the purification method described here would be useful due to easy scale-up and simple process without chromatographic process.     

Properties of recombinant 4-α-glucanotransferase from Bifidobacterium longum subsp. longum JCM 1217 and its application

Food Science and Biotechnology - To determine the physiochemical properties of the 4-α-glucanotransferase from Bifidobacterium sp., the bllj_0114 gene encoding 4-α-glucanotransferase was...     

Physicochemical properties and freeze–thaw stability of rice flour blends among rice cultivars with different amylose contents

Food Science and Biotechnology - The effectiveness of the rice flour blends (RFB) for improving the processing suitability of Dodamssal rice flour (DD), a functional rice variety with a relatively...     

Complex formation of a 4-α-glucanotransferase using starch as a biocatalyst for starch modification

Food Science and Biotechnology - A 4-α-glucanotransferases from Thermus thermophilus (TTαGT) possesses an extra substrate binding site, leading to facile purification of the intact enzyme...     

Rheological properties and permeability of soy protein-stabilised emulsion gels made by acidification with glucono-δ-lactone

BACKGROUND: Soy protein, an important efficient emulsifier, is widely used by the food industry for incorporation into milk, yogurts, ice cream, salad dressings, dessert products, etc. The objective of this study was to investigate the rheological and physical properties of soy protein‐stabilised emulsion gels as affected by protein concentration and gelation temperature. RESULTS: The rheological properties and permeability were determined using oscillatory rheometry, permeability and whey separation. The modulus (G′ and G″), fracture stress and fracture strain of acid‐induced emulsion gels after 20 h of glucono‐δ‐lactone addition depended strongly on soy protein concentration and gelation temperature. At increasing soy protein concentrations, acid‐induced emulsion gels had shorter gelation times but higher storage moduli (G′), fracture stresses and strains. Increasing gelation temperature decreased the gelation time, G′, fracture stresses and strains. Permeability and whey separation were significantly affected by the protein concentration and the gelation temperature. A significant positive correlation was observed between whey separation and permeability coefficient in emulsion gels formed at different temperatures. CONCLUSION: The rheological properties and permeability of soy protein‐stabilised emulsion gels were significantly influenced by protein concentration and gelation temperature. Copyright © 2011 Society of Chemical Industry     

Edible films made from blends of manioc starch and gelatin – Influence of different types of plasticizer and different levels of macromolecules on their properties

The interest in the development of edible and biodegradable films has increased because it is every day more evident that non-degradable materials are doing much damage to the environment. In this research, bioplastics were based on blends of manioc starch (native and modified) and gelatin in different proportions, added of glycerol or sorbitol, which were used as plasticizers. The objective was to study the effect of two different plasticizers, glycerol and sorbitol, and different concentrations of starch and gelatin on the barrier (water vapor permeability – WVP), mechanical (tensile strength and elongation at break), physicochemical (solubility in water and in acid) and physical properties (opacity and thickness) of the obtained bioplastics samples. As a result, all of them showed transparency and resistance to tensile strength, as well as increasing in thickness values and in the WVP, as the gelatin content increased in the formulations. Finally, all results for tensile strength and elongation at break obtained for those samples plasticized with sorbitol were better than those plasticized with glycerol.     

High-yield cycloamylose production from sweet potato starch using Pseudomonas isoamylase and Thermus aquaticus 4-α-glucanotransferase

Food Science and Biotechnology - An optimal reaction condition for producing cycloamylose (CA) from sweet potato starch was investigated using a combination of isoamylase (from Pseudomonas sp.) and...     

Study on the relationship between the degradation degrees of glutinous rice starch extruded with different α-amylases and the qualities of Chinese rice wine

In view that enzymatic extrusion can change the degradation degree of raw materials, this paper studied the regulation rules of four indexes of degradation degree of enzymatically extruded glutinous rice flour and the correlation between these indexes and the quality indexes of brewed Chinese rice wine. The results showed that at appropriate water content, dextrose equivalent (DE) and water solubility index (WSI) were linearly related to the enzyme content in extrusion with thermostable α-amylase (TS-αA) and mesophilic α-amylase (MS-αA), respectively. But water absorption index (WAI) was not significantly affected by enzyme content. The percentages of each degree of polymerisation (DP) (1–7) and that of the sum of the rest DPs called other DP were the fundamental reasons for different DE values, WSI, WAI and quality indexes of Chinese rice wine. DP1 was significantly positively correlated with alcohol degree, and negatively correlated with total acid. DP2–DP7 (except DP4) was significantly positively correlated with amino acid nitrogen. Besides, other DP was significantly positively correlated with total sugar and total acid. The percentages of DP1, DP2–DP7 and other DP could be respectively adjusted by changing enzyme content in extrusion with MS-αA, MS-αA/TS-αA and TS-αA to control the related quality indexes of Chinese rice wine.     

Molecular weight and structure of water soluble (1→3), (1→4)-β-glucans affect pasting properties of oat flours

Seven experimental oat lines with high (5.9% to 7.2%), medium (5.3% to 5.5%), and low (4.4%) β-glucan concentrations were evaluated for the effects of β-glucan molecular weight (MW) and structure on viscosities of oat-flour slurries. The MW of β-glucans was determined by size-exclusion high-performance liquid chromatography. The structural features of β-glucans were measured by using fluorophore-assisted capillary-electrophoresis after complete hydrolysis with lichenase. The oat-slurry viscosities were measured on a Rapid Visco Analyser under 4 conditions: (1) without starch (amylolysis, removal of starch by α-amylase); (2) without β-glucan (removal of β-glucan by lichenase); (3) natural action of enzymes (autolysis, in sodium buffer); and (4) inhibition of enzymes (in silver nitrate solution). Excluding one line (regression outlier), significant correlations (P < 0.05) between peak MW of β-glucan and viscosities of oat slurries were obtained under inhibition. The ratio of degree of polymerization (DP) 3/DP4 negatively correlated with viscosity under amylolysis, autolysis, and inhibition (P < 0.05). The amount of DP ≥ 5 negatively correlated with pasting final viscosity after β-glucan removal by lichenase (P < 0.05). Positive correlations (P < 0.05) between the ratio of β-(1→4)/β-(1→3) linkages and viscosities under autolysis and inhibition were found. Overall, these findings demonstrated that the peak MW, ratio of DP3/DP4, amount of DP ≥ 5, and ratio of β-(1→4)/β-(1→3) linkages of β-glucans impacted pasting properties of oat-flour slurries.     

Interaction of phenolic compounds with bovine serum albumin (BSA) and α-amylase and their relationship to astringency perception

The ability of grape seed extracts to bind to bovine serum albumin (BSA) and α-amylase was studied by fluorescence quenching of protein intrinsic fluorescence and nephelometry. The influence of grape seed ripeness on astringency was also evaluated. From the spectra obtained, the modified Sterm-Volmer (K(app)) and the bimolecular quenching constants were calculated. Results showed that grape seed extracts had good affinity for proteins. The association strength of tannin-protein interactions varied with changes in tannin structure associated with the degree of ripeness affecting the binding/quenching process. In all cases studied, higher values of K(app) were obtained in samples at harvest which have greater ability to bind to proteins than have samples at post-veraison time. Nephelometric assays show the same trend as do fluorescence quenching studies. A possible explanation for this is that, as seeds ripen, their tannins increase in molecular mass, which relates to an increase in hydrophobicity of the molecules, and this increases protein affinity. However, that is contrary to the reported decrease in astringency of grape seeds during maturity. This indicates that tannin-protein interactions are not the only explanation for the complex sensations of astringency of grape seeds.     

Use of micellar casein concentrate and milk protein concentrate treated with transglutaminase in imitation cheese products—Melt and stretch properties

Melt and stretch properties in dairy-based imitation mozzarella cheese (IMC) are affected by the amount of intact casein provided by dairy ingredients in the formulation. Rennet casein (RCN) is the preferred ingredient to provide intact casein in a formulation. Ingredients produced using membrane technology, such as milk protein concentrate (MPC) and micellar casein concentrate (MCC), are unable to provide the required functionality. However, the use of transglutaminase (TGase) has potential to modify the physical properties of MPC or MCC and may improve their functionality in IMC. The objective of this study was to determine the effect of TGase-treated MPC and MCC retentates on melt and stretch properties when they are used in IMC and to compare them with IMC made using RCN. The MCC and MPC retentates were produced using 3 different lots of pasteurized skim milk and treated with 3 levels of TGase enzyme: no TGase (control), low TGase: 0.3 units/g of protein, and high TGase: 3.0 units/g of protein. Each of the MCC and MPC treatments was heated to 72°C for 10 min to inactivate TGase and then spray dried. Each MCC, MPC, and RCN powder was then used in an IMC formulation that was standardized to 48% moisture, 21% fat, 20% protein, and 1% salt. The IMC were manufactured in a twin-screw cooker by blending, mixing, and heating various ingredients (4.0 kg). Due to extensive crosslinking, the IMC formulation with the highest TGase level (MCC or MPC) did not form an emulsion. The IMC made from MCC treatments had significantly higher stretchability on pizza compared with their respective MPC treatments. The IMC made from TGase-treated MCC and MPC had significantly lower melt area and significantly higher transition temperature (TT) and stretchability compared with their respective controls. Comparison of IMC made using TGase-treated MCC and MPC to the RCN IMC indicated no difference in TT or texture profile analysis-stretchability; however, the Schreiber melt test area was significantly lower. Our results demonstrated that TGase treatment modifies the melt and stretch characteristics of MCC and MPC in IMC applications, and TGase-treated MPC and MCC can be used to replace RCN in IMC formulations.     

Evaluation of luteolysis,follicle size,and time to ovulation in Holstein heifers treated with two different analogs and doses of prostaglandin-F2α

Objectives were to evaluate the effect of 2 analogs of PGF_2α (cloprostenol vs. dinoprost) and 2 doses (1 injection vs. 2 injections) on luteolysis, follicle diameter, hormonal concentrations, and time to ovulation in dairy heifers. Holstein heifers were fitted with automated estrus detection devices and had their estrous cycle synchronized using PGF_2α and an intravaginal insert containing progesterone. Heifers detected in estrus were blocked by weight and randomly assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement: cloprostenol on d 7 after estrus (CLOx1; n = 45), cloprostenol on d 7 and 8 after estrus (CLOx2; n = 41), dinoprost on d 7 after estrus (DINx1; n = 43), or dinoprost on d 7 and 8 after estrus (DINx2; n = 44). Treatment with the first injection of PGF_2α was defined as experiment d 0. Area and blood flow of corpus luteum (CL) and diameter of follicles >5 mm were recorded every 12 h from d 0 to estrus and every 6 h thereafter until ovulation. Blood was sampled every 6 h from d 0 until ovulation. Heifers treated with cloprostenol had shorter interval to luteolysis (± SEM; CLOx1 = 23.5 ± 2.2, CLOx2 = 22.9 ± 2.2, DINx1 = 32.6 ± 2.7, DINx2 = 26.4 ± 2.1 h); however, time to ovulation was not affected by treatment. A smaller proportion of heifers treated with a single injection of PGF_2α underwent luteolysis compared with heifers treated with 2 injections (CLOx1 = 84.6 ± 6.2, CLOx2 = 100.0 ± 0.0, DINx1 = 59.7 ± 9.8, DINx2 = 96.3 ± 2.7%). Proportion of heifers that ovulated was smaller for DINx1 compared with other treatments (CLOx1 = 88.8 ± 5.1, CLOx2 = 100.0 ± 0.0, DINx1 = 55.2 ± 9.7, DINx2 = 94.4 ± 3.4%). Ovulatory follicle diameter was larger for DINx1 (18.2 ± 2.7 mm) compared with DINx2 (17.4 ± 2.7 mm), whereas dose did not affect the diameter of the ovulatory follicle in heifers treated with cloprostenol (CLOx1 = 17.6 ± 2.7 vs. CLOx2 = 17.8 ± 2.8 mm). Among heifers that underwent luteolysis, progesterone concentrations from 18 to 36 h after treatment were lesser in heifers treated with cloprostenol compared with those treated with dinoprost. Type of PGF_2α did not affect progesterone concentrations past 36 h from treatment; however, heifers treated with 2 PGF_2α injections had lesser progesterone concentrations and CL blood flow from 36 to 72 h after treatment compared with heifers that received a single PGF_2α injection.     

Viability of Listeria monocytogenes on commercially-prepared hams surface treated with acidic calcium sulfate and lauric arginate and stored at 4°C

We demonstrated the effectiveness of delivering an antimicrobial purge/fluid into shrink-wrap bags immediately prior to introducing the product and vacuum sealing, namely the “Sprayed Lethality In Container” (SLIC™) intervention delivery method. The pathogen was Listeria monocytogenes, the antimicrobials were acidic calcium sulfate (ACS; calcium sulfate plus lactic acid; 1:1 or 1:2 in dH₂O) and lauric arginate (LAE; Ethyl-N-dodecanoyl-l-arginate hydrochloride; 5% or 10% in dH₂O), and the product was commercially prepared “table brown” ham (ca. 3 pounds each). Hams were surface inoculated with a five-strain cocktail of L. monocytogenes (ca. 7.0 log₁₀ CFU per ham), added to shrink-wrap bags that already contained ACS or LAE, vacuum-sealed, and stored at 4 °C for 24 h. Pathogen levels decreased by 1.2, 1.6, 2.4, and 3.1 log₁₀ CFU/ham and 0.7, 1.6, 2.2, and 2.6 log₁₀ CFU/ham in samples treated with 2, 4, 6, and 8 mL of a 1:1 and 1:2 solution of ACS, respectively. In samples treated with 2, 4, 6, and 8 mL of a 5% solution of LAE, pathogen levels decreased by 3.3, 6.5, 5.6, and 6.5 log₁₀ CFU/ham, whereas when treated with a 10% solution of LAE pathogen levels decreased ca. 6.5 log₁₀ CFU/ham for all application volumes tested. The efficacy of ACS and LAE were further evaluated in shelf-life studies wherein hams were surface inoculated with either ca. 3.0 or 7.0 log₁₀ CFU of L. monocytogenes, added to shrink-wrap bags that contained 0, 4, 6, or 8 mL of either a 1:2 solution of ACS or a 5% solution of LAE, vacuum-sealed, and stored at 4 °C for 60 days. For hams inoculated with 7.0 log₁₀ CFU, L. monocytogenes levels decreased by ca.1.2, 1.5, and 2.0 log₁₀ CFU/ham and 5.1, 5.4, and 5.5 log₁₀ CFU/ham within 24 h at 4 °C in samples treated with 4, 6, and 8 mL of a 1:2 solution of ACS and a 5% solution of LAE, respectively, compared to control hams that were not treated with either antimicrobial. Thereafter, pathogen levels remained relatively unchanged (±1.0 log₁₀ CFU/ham ) after 60 days at 4 °C in hams treated with 4, 6, and 8 mL of a 1:2 solution of ACS and increased by ca. 2.0–5.0 log₁₀ CFU/ham in samples treated with 4, 6, and 8 mL of a 5% solution of LAE. For hams inoculated with 3.0 log₁₀ CFU, L. monocytogenes levels decreased by 1.3, 1.9, and 1.8 log₁₀ CFU/ham within 24 h at 4 °C in samples treated with 4, 6, and 8 mL of a 1:2 solution of ACS, respectively, compared to control hams that were not treated. Likewise, levels of the pathogen were reduced to below the limit of detection (i.e., 1.48 log₁₀ CFU/ham) in the presence of 4, 6, and 8 mL of a 5% solution of LAE within 24 h at 4 °C. After 60 days at 4 °C, pathogen levels remained relatively unchanged (±0.3 log₁₀ CFU/ham) in hams treated with 4, 6, and 8 mL of a 1:2 solution of ACS. However, levels of L. monocytogenes increased by ca. 2.0 log₁₀ CFU/ham in samples treated with 4 and 6 mL of a 5% LAE solution within 60 days but remained below the detection limit on samples treated with 8 mL of this antimicrobial. These data confirmed that application via SLIC™ of both ACS and LAE, at the concentrations and volumes used in this study, appreciably reduced levels of L. monocytogenes on the surface of hams within 24 h at 4 °C and showed potential for controlling outgrowth of the pathogen over 60 days of refrigerated storage.     

Effects of turmeric, shallot extracts, and their combination on quality characteristics of vacuum-packaged rainbow trout stored at 4 ± 1 °C

The effects of turmeric extract (T), shallot extract (Sh), and their combination (T + Sh) on the quality of vacuum-packaged rainbow trout (Oncorhynchus mykiss) were examined during refrigerated storage (4 ± 1 °C) over a period of 20 d. Fish samples were divided into 4 batches; 3 batches were treated by dipping for 30 min in aqueous solution of turmeric extract (1.5%; v/v), shallot extract (1.5%; v/v), or turmeric and shallot extract combination (1.5%+ 1.5% v/v), while the fourth batch was dipped in distilled water as a control sample. The control and the treated fish samples were analyzed periodically for microbiological (total viable count, psychrotrophic count), chemical (total volatile base nitrogen [TVB-N], peroxide value [PV], and thiobarbituric acid [TBA] value), and sensory characteristics. The results indicated that the effect of the T, Sh, and T + Sh on the fish samples were to enable the good quality characteristics to be retained longer and to extend the shelf life during the refrigerated storage.     

Rheological behaviors,structural properties and freeze–thaw stability of normal and waxy genotypes of barley starch: a comparative study with mung bean,potato, and corn starches

Food Science and Biotechnology - The rheological behaviors, structural properties and freeze-thaw stability of starch isolated from Tetonia barley (Normal genotype, Reg. No. CV-334, PI 646199) and...     

Spores of Bacillus cereus strain KBAB4 produced at 10 °C and 30 °C display variations in their properties

Spores of the psychrotrophic Bacillus cereus KBAB4 strain were produced at 10 °C and 30 °C in fermentors. Spores produced at 30 °C were more resistant to wet heat at 85 °C, 1 % glutaraldehyde, 5 % hydrogen peroxide, 1 M NaOH and pulsed light at fluences between 0.5 and 1.75 J cm⁻² and to a lesser extent to monochromatic UV-C at 254 nm. No difference in resistance to 0.25 mM formaldehyde, 1 M nitrous acid and 0.025 g l⁻¹ calcium hypochlorite was observed. Spores produced at 10 °C germinated more efficiently with 10 mM and 100 mM l-alanine than spores produced at 30 °C, while no difference in germination was observed with inosine. Dipicolinic acid (DPA) content in the spore was significantly higher for spores prepared at 30 °C. Composition of certain fatty acids varied significantly between spores produced at 10 °C and 30 °C.