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Fulltext Production of chitosan oligosaccharides using β-glycosidic degrading enzyme: optimization using response surface methodology

Yusof Nurhayati, Abdul Manaf Ali

Malaysian Journal of Applied Sciences, 2020;5(2):30-44.
MyJurnal

Many researchers have focused chitosan as a source of potential bioactive material during the past few decades. However, chitosan has several drawbacks to be utilised in biological applications, including poor solubility under physiological conditions. Therefore, a new interest has recently emerged on partially hydrolysed chitosan, chitosan oligosaccharides (COS). In this study, degradation of chitosan was performed by Cellulase from Trichoderma reesei® 1.5L and Response Surface Methodology (RSM) were employed to optimize the hydrolysis temperature, pH, enzyme concentration and substrate concentration. Optimization of cellulase T. reesei® using central composite design (CCD) was to obtain optimum parameters and all the factors showed significant effects (p˂0.05). The maximum response, Celluclast® activity (1.268 U) was obtained by assaying the process at 49.79oC, pH 4.5, 3% (v/w) of enzyme concentration and 25% (w/v) concentration of chitosan for 24 hours.

Matched MeSH terms: Oligosaccharides
Synergistic effects of dietary supplementation of Bacillus subtilis WB60 and mannanoligosaccharide (MOS) on growth performance, immunity and disease resistance in Japanese eel, Anguilla japonica

Lee S, Katya K, Hamidoghli A, Hong J, Kim DJ, Bai SC

Fish Shellfish Immunol, 2018 Dec;83:283-291.
PMID: 30217508 DOI: 10.1016/j.fsi.2018.09.031

This study evaluated the synergistic effects of dietary Bacillus subtilis WB60 and mannanoligosaccharide (MOS) in juvenile Japanese eel, Anguilla japonica. Seven treatment diets were formulated to contain three different levels of B. subtilis (0.0, 0.5, and 1.0 × 107 CFU/g diet denoted as BS0, BS0.5, and BS1, respectively) with two MOS levels (0 and 5 g/kg diet denoted as M0 and M5, respectively), and one diet with oxytetracycline (OTC) at 5 g/kg diet. Each diet (BS0M0 (CON), BS0M5, BS0.5M0, BS0.5M5, BS1M0, BS1M5, and OTC) was fed to triplicate groups of 20 fish averaging 9.00 ± 0.11 g (mean ± SD) for eight weeks. Average weight gain, feed efficiency, specific growth rate and protein efficiency ratio of fish fed the BS0.5M5 and BS1M5 diets were significantly higher than those of fish fed CON, BS0.5M0 and OTC diets (P  0.05). Therefore, the results for growth performance, non-specific immune responses, intestinal morphology, and disease resistance demonstrated that supplementation of B. subtilis at 0.5 × 107 CFU/g diet and mannanoligosaccharide at 5 g/kg diet could have beneficial synergistic effects in Japanese eel. The isolated probiotic from eel and the selected prebiotic could lead to the development of a specific and potential synbiotic in Japanese eel aquaculture.

Matched MeSH terms: Oligosaccharides/pharmacology*
Valorization of renewable resources to functional oligosaccharides: Recent trends and future prospective

Narisetty V, Parhi P, Mohan B, Hakkim Hazeena S, Naresh Kumar A, Gullón B, et al.

Bioresour Technol, 2022 Feb;346:126590.
PMID: 34953996 DOI: 10.1016/j.biortech.2021.126590

Lignocellulosic wastes have the ability to be transformed into oligosaccharides and other value-added products. The synthesis of oligosaccharides from renewable sources bestow to growing bioeconomies. Oligosaccharides are synthesized chemically or biologically from agricultural residues. These oligosaccharides are functional food supplements that have a positive impact on humans and livestock. Non-digestible oligosaccharides, refered as prebiotics are beneficial for the colonic microbiota inhabiting the f the digestive system. These microbiota plays a crucial role in stimulating the host immune system and other physiological responses. The commonly known prebiotics, galactooligosaccharides (GOS), xylooligosaccharides (XOS), fructooligosaccharides (FOS), mannanooligosaccharides (MOS), and isomaltooligosaccharides (IOS) are synthesized either through enzymatic or whole cell-mediated approaches using natural or agricultural waste substrates. This review focusses on recent advancements in biological processes, for the synthesis of oligosaccharides using renewable resources (lignocellulosic substrates) for sustainable circular bioeconomy. The work also addresses the limitations associated with the processes and commercialization of the products.

Matched MeSH terms: Oligosaccharides*
Fulltext Sensory descriptive analysis and consumer acceptability of original “kaya” and “kaya” partially substituted with inulin

Phang, Y.L., Chan, H.K.

International Food Research Journal, 2009;16(4):483-492.
MyJurnal

A study was done on the feasibility of inulin, a fructooligosaccharides with the health benefits of prebiotic, as partial sugar replacer in “kaya” by investigating the effects of different substitution levels (0%; control, 10%, 30% and 50%) on the sensory properties. The samples were investigated by 10 descriptive panelists for colour, sweetness, smoothness, firmness, adhesiveness and spreadability attributes before further evaluated by 100 consumers for acceptance levels. Consistent with the descriptive panel, the evaluation by the consumers showed formulated samples at 10% and 30% were comparable to the commercial “kaya” being no significant difference (p>0.05) found in acceptability scores. 50% substitution received lower acceptability scores and was significantly different (p

Matched MeSH terms: Oligosaccharides
Purification and characterization of 31-kDa palm pollen glycoprotein (Ela g Bd 31 K), which is recognized by IgE from palm pollinosis patients

Kimura Y, Maeda M, Kimupa M, Lai OM, Tan SH, Hon SM, et al.

Biosci Biotechnol Biochem, 2002 Apr;66(4):820-7.
PMID: 12036055

A basic glycoprotein, which was recognized by IgE from oil palm pollinosis patients, has been purified from oil palm pollen (Elaeis guineensis Jacq.), which is a strong allergen and causes severe pollinosis in Malaysia and Singapore. Soluble proteins were extracted from defatted palm pollen with both Tris-HCl buffer (pH 7.8) and Na-acetate buffer (pH 4.0). The allergenic glycoprotein was purified from the total extract to homogeneity with 0.4% yield by a combination of DEAE- and CM-cellulose, SP-HPLC, and gel filtration. The purified oil palm pollen glycoprotein with molecular mass of 31 kDa was recognized by the beta1-2 xylose specific antibody, suggesting this basic glycoprotein bears plant complex type N-glycan(s). The palm pollen basic glycoprotein, designated Ela g Bd 31 K, was recognized by IgE of palm pollinosis patients, suggesting Ela g Bd 31 K should be one of the palm pollen allergens. The preliminary structural analysis of N-glycans linked to glycoproteins of palm pollens showed that the antigenic N-glycans having alpha1-3 fucose and alpha1-2 xylose residues (GlcNAc(2 to approximately 0)Man3Xyl1Fuc(1 to approximately 0)GlcNAc2) actually occur on the palm pollen glycoproteins, in addition to the high-mannose type structures (Man(9 to approximately 5)GlcNAc2).

Matched MeSH terms: Oligosaccharides/chemistry*
Fulltext Efficacy of microbial cell preparation in improving chronic constipation: a randomized, double-blind, placebo-controlled trial

Jayasimhan S, Yap NY, Roest Y, Rajandram R, Chin KF

Clin Nutr, 2013 Dec;32(6):928-34.
PMID: 23561636 DOI: 10.1016/j.clnu.2013.03.004

Probiotics is an emerging therapeutic agent which may alleviate the symptoms of constipation. We evaluated the effectiveness of microbial cell preparation (Hexbio(®)) containing fructooligosaccharide, Bifidobacterium and Lactobacillus in improving stool frequency and symptoms of chronic constipation.

Matched MeSH terms: Oligosaccharides/administration & dosage
Fulltext Modified bicinchoninic acid assay for accurate determination of variable length reducing sugars in carbohydrates

Hussain, H., Ngaini, Z., Chong, N.F-M.

International Food Research Journal, 2018;25(6):2614-2619.
MyJurnal

The accurate determination of reducing ends of malto-oligosaccharides is essential for calculating the enzyme activities of starch debranching enzymes. The suitability of the 3,5-Dinitrosalicylic acid (DNS) method, the Dygert method, and the Bicinchoninic acid (BCA) method for accurate determination of reducing ends from malto-oligosaccharides of different chain lengths is compared. The results showed that BCA assay was much more accurate than the other assays. The results for the BCA assay showed that different malto-oligosaccharides gave observed (measured) values that were significantly similar to the expected (predetermined) values. In contrast, the DNS and Dygert assays underestimated the amount of reducing sugar present for glucose. Furthermore, both DNS and Dygert methods showed increasing degree of overestimation of the amount of reducing sugar present with the increasing length of the malto-oligosaccharide sugar chains. The BCA assay can suitably quantify reducing sugars even in mixtures of oligosaccharides with different chain lengths. Thus, enzyme activities can be measured without bias towards higher values for enzymes that preferentially cleave the longer chain lengths.

Matched MeSH terms: Oligosaccharides
Use of the glucose analyser in the estimation of small bowel mucosal oligosaccharidases

Iyngkaran, N., Yadav, M., Boey, G.B., Davis, K.

Malaysian Journal of Child Health, 1991;3(1):37-39.
MyJurnal

Estimation of oligosaccharidases in the jejunal mucosa is useful in the diagnosis and evaluation of primary and secondary oligosaccharide intolerance. Until recently these enzymes have been estimated by the method of Dahlgvist.4While the method is accurate and reliable it is tedious and cumbersome. We describe here a semi quantitative method, using the glucose analyser. (Copied from article).

Matched MeSH terms: Oligosaccharides
Fulltext Human Milk Oligosaccharide, Phospholipid, and Ganglioside Concentrations in Breast Milk from United Arab Emirates Mothers: Results from the MISC Cohort

McJarrow P, Radwan H, Ma L, MacGibbon AKH, Hashim M, Hasan H, et al.

Nutrients, 2019 Oct 08;11(10).
PMID: 31597293 DOI: 10.3390/nu11102400

Human milk oligosaccharides (HMOs), phospholipids (PLs), and gangliosides (GAs) are components of human breast milk that play important roles in the development of the rapidly growing infant. The differences in these components in human milk from the United Arab Emirates (UAE) were studied in a cross-sectional trial. High-performance liquid chromatography‒mass spectrometry was used to determine HMO, PL, and GA concentrations in transitional (5-15 days) and mature (at 6 months post-partum) breast milk of mothers of the United Arab Emirates (UAE). The results showed that the average HMO (12 species), PL (7 species), and GA (2 species) concentrations quantified in the UAE mothers' transitional milk samples were (in mg/L) 8204 ± 2389, 269 ± 89, and 21.18 ± 11.46, respectively, while in mature milk, the respective concentrations were (in mg/L) 3905 ± 1466, 220 ± 85, and 20.18 ± 9.75. The individual HMO concentrations measured in this study were all significantly higher in transitional milk than in mature milk, except for 3 fucosyllactose, which was higher in mature milk. In this study, secretor and non-secretor phenotype mothers showed no significant difference in the total HMO concentration. For the PL and GA components, changes in the individual PL and GA species distribution was observed between transitional milk and mature milk. However, the changes were within the ranges found in human milk from other regions.

Matched MeSH terms: Oligosaccharides/analysis*
Fulltext The impact of oligofructose on stimulation of gut hormones, appetite regulation and adiposity

Daud NM, Ismail NA, Thomas EL, Fitzpatrick JA, Bell JD, Swann JR, et al.

Obesity (Silver Spring), 2014 Jun;22(6):1430-8.
PMID: 24715424 DOI: 10.1002/oby.20754

OBJECTIVE: To investigate the effect of nutrient stimulation of gut hormones by oligofructose supplementation on appetite, energy intake (EI), body weight (BW) and adiposity in overweight and obese volunteers.
METHODS: In a parallel, single-blind and placebo-controlled study, 22 healthy overweight and obese volunteers were randomly allocated to receive 30 g day(-1) oligofructose or cellulose for 6 weeks following a 2-week run-in. Subjective appetite and side effect scores, breath hydrogen, serum short chain fatty acids (SCFAs), plasma gut hormones, glucose and insulin concentrations, EI, BW and adiposity were quantified at baseline and post-supplementation.
RESULTS: Oligofructose increased breath hydrogen (P

Matched MeSH terms: Oligosaccharides/administration & dosage*
Characterization of a type I pullulanase from Anoxybacillus sp. SK3-4 reveals an unusual substrate hydrolysis

Kahar UM, Ng CL, Chan KG, Goh KM

Appl Microbiol Biotechnol, 2016 Jul;100(14):6291-307.
PMID: 27000839 DOI: 10.1007/s00253-016-7451-6

Type I pullulanases are enzymes that specifically hydrolyse α-1,6 linkages in polysaccharides. This study reports the analyses of a novel type I pullulanase (PulASK) from Anoxybacillus sp. SK3-4. Purified PulASK (molecular mass of 80 kDa) was stable at pH 5.0-6.0 and was most active at pH 6.0. The optimum temperature for PulASK was 60 °C, and the enzyme was reasonably stable at this temperature. Pullulan was the preferred substrate for PulASK, with 89.90 % adsorbance efficiency (various other starches, 56.26-72.93 % efficiency). Similar to other type I pullulanases, maltotriose was formed on digestion of pullulan by PulASK. PulASK also reacted with β-limit dextrin, a sugar rich in short branches, and formed maltotriose, maltotetraose and maltopentaose. Nevertheless, PulASK was found to preferably debranch long branches at α-1,6 glycosidic bonds of starch, producing amylose, linear or branched oligosaccharides, but was nonreactive against short branches; thus, no reducing sugars were detected. This is surprising as all currently known type I pullulanases produce reducing sugars (predominantly maltotriose) on digesting starch. The closest homologue of PulASK (95 % identity) is a type I pullulanase from Anoxybacillus sp. LM14-2 (Pul-LM14-2), which is capable of forming reducing sugars from starch. With rational design, amino acids 362-370 of PulASK were replaced with the corresponding sequence of Pul-LM14-2. The mutant enzyme formed reducing sugars on digesting starch. Thus, we identified a novel motif involved in substrate specificity in type I pullulanases. Our characterization may pave the way for the industrial application of this unique enzyme.

Matched MeSH terms: Oligosaccharides/chemistry
Low FODMAP diet significantly improves IBS symptoms: an Irish retrospective cohort study

Nawawi KNM, Belov M, Goulding C

Eur J Nutr, 2020 Aug;59(5):2237-2248.
PMID: 31520160 DOI: 10.1007/s00394-019-02074-6

INTRODUCTION: There is growing evidence that a diet low in fermentable oligosaccharides, disaccharides, monosaccharides, and polyols (FODMAP) improves symptoms in irritable bowel syndrome (IBS) patients. We aimed to retrospectively investigate the effects of this diet in Irish IBS cohort over a 12-month follow-up period, including after re-introduction of the high FODMAP foods.
METHODS: All the tertiary referrals seen by an FODMAP-trained dietician were reviewed (2013-2016). Patients were evaluated for IBS symptoms by a questionnaire (four-point Likert scale). Subsequently, advice regarding the low FODMAP diet was given. Symptoms' response was assessed at 3-, 6-, and 12-month follow-up, by use of the same questionnaire. Re-introduction of high FODMAP foods was aimed to commence at the subsequent follow-up.
RESULTS: A total of 164 patients were identified. Thirty-seven patients were excluded due to failure to attend for follow-up. Hundred and twenty-seven patients (77% patients, of which 85% were female) completed the initial 3-month follow-up. Forty-five percent (74/164) and twenty-five percent (41/164) of the patients had continued follow-up at 6 and 12 months, respectively. Of the 127 patients who returned for follow-up, their commonest baseline symptoms were lethargy (92%), bloating (91%), flatulence (91%), and abdominal pain (89%). All symptoms were significantly improved at the initial follow-up (p

Matched MeSH terms: Oligosaccharides
Fulltext Phytochemical Properties and Nutrigenomic Implications of Yacon as a Potential Source of Prebiotic: Current Evidence and Future Directions

Cao Y, Ma ZF, Zhang H, Jin Y, Zhang Y, Hayford F

Foods, 2018 Apr 12;7(4).
PMID: 29649123 DOI: 10.3390/foods7040059

The human gut is densely populated with diverse microbial communities that are essential to health. Prebiotics and fiber have been shown to possess the ability to modulate the gut microbiota. One of the plants being considered as a potential source of prebiotic is yacon. Yacon is an underutilized plant consumed as a traditional root-based fruit in South America. Yacon mainly contains fructooligosaccharides (FOS) and inulin. Therefore, it has bifidogenic benefits for gut health, because FOS are not easily broken down by digestive enzymes. Bioactive chemical compounds and extracts isolated from yacon have been studied for their various nutrigenomic properties, including as a prebiotic for intestinal health and their antimicrobial and antioxidant effects. This article reviewed scientific studies regarding the bioactive chemical compounds and nutrigenomic properties of extracts and isolated compounds from yacon. These findings may help in further research to investigate yacon-based nutritional products. Yacon can be considered a potential prebiotic source and a novel functional food. However, more detailed epidemiological, animal, and human clinical studies, particularly mechanism-based and phytopharmacological studies, are lacking for the development of evidence-based functional food products.

Matched MeSH terms: Oligosaccharides
Effects of dietary prebiotics, probiotic and synbiotics on performance, caecal bacterial populations and caecal fermentation concentrations of broiler chickens

Mookiah S, Sieo CC, Ramasamy K, Abdullah N, Ho YW

J Sci Food Agric, 2014 Jan 30;94(2):341-8.
PMID: 24037967 DOI: 10.1002/jsfa.6365

In view of a worldwide attempt to restrict or ban the use of antibiotics as growth promoters in animal production, probiotics, prebiotics and combinations of both, as synbiotics, have been suggested as potential alternatives. In this study, the effects of a prebiotic (isomalto-oligosaccharides, IMO), a multi-strain probiotic (consisting of 11 Lactobacillus strains), and a combination of these dietary additives as a synbiotic on the performance, caecal bacterial populations and concentrations of caecal volatile fatty acids and non-volatile fatty acids of broiler chickens were evaluated.

Matched MeSH terms: Oligosaccharides/pharmacology*
Functional characterisation and product specificity of Endo-β-1,3-glucanase from alkalophilic bacterium, Bacillus lehensis G1

Jaafar NR, Khoiri NM, Ismail NF, Mahmood NAN, Abdul Murad AM, Abu Bakar FD, et al.

Enzyme Microb Technol, 2020 Oct;140:109625.
PMID: 32912685 DOI: 10.1016/j.enzmictec.2020.109625

Endo-β-1,3-glucanase from alkalophilic bacterium, Bacillus lehensis G1 (Blg32) composed of 284 amino acids with a predicted molecular mass of 31.6 kDa is expressed in Escherichia coli and purified to homogeneity. Herein, Blg32 characteristics, substrates and product specificity as well as structural traits that might be involved in the production of sugar molecules are analysed. This enzyme functions optimally at the temperature of 70 °C, pH value of 8.0 with its catalytic activity strongly enhanced by Mn2+. Remarkably, the purified enzyme is highly stable in high temperature and alkaline conditions. It exhibits the highest activity on laminarin (376.73 U/mg) followed by curdlan and yeast β-glucan. Blg32 activity increased by 62% towards soluble substrate (laminarin) compared to insoluble substrate (curdlan). Hydrolytic products of laminarin were oligosaccharides with degree of polymerisation (DP) of 1 to 5 with the main product being laminaritriose (DP3). This suggests that the active site of Blg32 could recognise up to five glucose units. High concentration of Blg32 mainly produces glucose whilst low concentration of Blg32 yields oligosaccharides with different DP (predominantly DP3). A theoretical structural model of Blg32 was constructed and structural analysis revealed that Trp156 is involved in multiple hydrophobic stacking interactions. The amino acid was predicted to participate in substrate recognition and binding. It was also exhibited that catalytic groove of Blg32 has a narrow angle, thus limiting the substrate binding reaction. All these properties and knowledge of the subsites are suggested to be related to the possible mode of action of how Blg32 produces glucooligosaccharides.

Matched MeSH terms: Oligosaccharides/metabolism
Entrapment of porous cross-linked enzyme aggregates of maltogenic amylase from Bacillus lehensis G1 into calcium alginate for maltooligosaccharides synthesis

Nawawi NN, Hashim Z, Rahman RA, Murad AMA, Bakar FDA, Illias RM

Int J Biol Macromol, 2020 May 01;150:80-89.
PMID: 32035147 DOI: 10.1016/j.ijbiomac.2020.02.032

Maltooligosaccharides (MOSs) are emerging oligosaccharides in food-based applications and can be synthesized through the enzymatic synthesis of maltogenic amylase from Bacillus lehensis G1 (Mag1). However, the lack of enzyme stability makes this approach unrealistic for industrial applications. The formation of cross-linked enzyme aggregates (CLEAs) is a promising tool for improving enzyme stability, and the substrate accessibility problem of CLEA formation was overcome by the addition of porous agents to generate porous CLEAs (p-CLEAs). However, p-CLEAs exhibited high enzyme leaching and low solvent tolerance. To address these problems, p-CLEAs of Mag1 (Mag1-p-CLEAs) were entrapped in calcium alginate beads (CA). Mag1-p-CLEAs-CA prepared with 2.5% (w/v) sodium alginate and 0.6% (w/v) calcium chloride yielded 53.16% (17.0 U/mg) activity and showed a lower deactivation rate and longer half-life than those of entrapped free Mag1 (Mag1-CA) and entrapped non-porous Mag1-CLEAs (Mag1-CLEAs-CA). Moreover, Mag1-p-CLEAs-CA exhibited low enzyme leaching and high tolerance in various solvents compared to Mag1-p-CLEAs. A kinetic study revealed that Mag1-p-CLEAs-CA exhibited relatively high affinity towards beta-cyclodextrin (β-CD) (Km = 0.62 mM). MOSs (300 mg/g) were synthesized by Mag1-p-CLEAs-CA at 50 °C. Finally, the reusability of Mag1-p-CLEAs-CA makes them as a potential biocatalyst for the continuous synthesis of MOSs.

Matched MeSH terms: Oligosaccharides/biosynthesis*; Oligosaccharides/chemistry
Combined cross-linked enzyme aggregates of cyclodextrin glucanotransferase and maltogenic amylase from Bacillus lehensis G1 for maltooligosaccharides synthesis

Yip YS, Manas NHA, Jaafar NR, Rahman RA, Puspaningsih NNT, Illias RM

Int J Biol Macromol, 2023 Jul 01;242(Pt 1):124675.
PMID: 37127056 DOI: 10.1016/j.ijbiomac.2023.124675

Maltooligosaccharides (MOS) are functional oligosaccharides that can be synthesized through enzymatic cascade reaction between cyclodextrin glucanotransferase (CGTase) and maltogenic amylase (Mag1) from Bacillus lehensis G1. To address the problems of low operational stability and non-reusability of free enzymes, both enzymes were co-immobilized as combined cross-linked enzyme aggregates (Combi-CLEAs-CM) with incorporation of bovine serum albumin (BSA) and Tween 80 (Combi-CLEAs-CM-add). Combi-CLEAs-CM and Combi-CLEAs-CM-add showed activity recoveries of 54.12 % and 69.44 %, respectively after optimization. Combi-CLEAs-CM-add showed higher thermal stability at higher temperatures (40 °C) with longer half-life (46.20 min) as compared to those of free enzymes (36.67 min) and Combi-CLEAs-CM (41.51 min). Both combi-CLEAs also exhibited higher pH stability over pH 5 to pH 9, and displayed excellent reusability with >50 % of initial activity retained after four cycles. The reduction in Km value of about 22.80 % and 1.76-fold increase in starch hydrolysis in comparison to Combi-CLEAs-CM attested the improvement of enzyme-substrate interaction by Tween 80 and pores formation by BSA in Combi-CLEAs-CM-add. The improved product specificity of Combi-CLEAs-CM-add also produced the highest yield of MOS (492 mg/g) after 3 h. Therefore, Combi-CLEAs-CM-add with ease of preparation, excellent reusability and high operational stability is believed to be highly efficacious biocatalyst for MOS production.

Matched MeSH terms: Oligosaccharides
Innovative biocatalyst synthesis of pectinolytic enzymes by cross-linking strategy: Potentially immobilised pectinases for the production of pectic-oligosaccharides from pectin

Abd Rahman NH, Rahman RA, Rahmat Z, Jaafar NR, Puspaningsih NNT, Illias RM

Int J Biol Macromol, 2024 Jan;256(Pt 1):128260.
PMID: 38000618 DOI: 10.1016/j.ijbiomac.2023.128260

Pectinases are outstanding multienzymes, which have the potential to produce new emerging pectic-oligosaccharides (POS) via enzymatic hydrolysis of pectin. However, free pectinase is unable to undergo repeated reaction for the production of POS. This study proposed a sustainable biocatalyst of pectinases known as cross-linked pectinase aggregates (CLPA). Pectinase from Aspergillus aculeatus was successfully precipitated using 2 mg/mL pectinase and 60 % acetone for 20 min at 20 °C, which remained 36.3 % of its initial activity. The prepared CLPA showed the highest activity recovery (85.0 %), under the optimised conditions (0.3 % (v/v) starch and glutaraldehyde mixture (St/Ga), 1.5: 1 of St/Ga, 25 °C, 1.5 h). Furthermore, pectin-degrading enzymes from various sources were used to produce different CLPA. The alteration of pectinase secondary structure gave high stability in acidic condition (pH 4), thermostability, deactivation energy and half-life, and improved storage stability at 4 °C for 30 days. Similarly to their free counterpart, the CLPA exhibited comparable enzymatic reaction kinetics and could be reused eight times with approximately 20 % of its initial activity. The developed CLPA does not only efficaciously produced POS from pectin as their free form, but also exhibited better operational stability and reusability, making it more suitable for POS production.

Matched MeSH terms: Oligosaccharides/chemistry
Fulltext The characterisation of an alkali-stable maltogenic amylase from Bacillus lehensis G1 and improved malto-oligosaccharide production by hydrolysis suppression

Abdul Manas NH, Pachelles S, Mahadi NM, Illias RM

PLoS One, 2014;9(9):e106481.
PMID: 25221964 DOI: 10.1371/journal.pone.0106481

A maltogenic amylase (MAG1) from alkaliphilic Bacillus lehensis G1 was cloned, expressed in Escherichia coli, purified and characterised for its hydrolysis and transglycosylation properties. The enzyme exhibited high stability at pH values from 7.0 to 10.0. The hydrolysis of β-cyclodextrin (β-CD) produced malto-oligosaccharides of various lengths. In addition to hydrolysis, MAG1 also demonstrated transglycosylation activity for the synthesis of longer malto-oligosaccharides. The thermodynamic equilibrium of the multiple reactions was shifted towards synthesis when the reaction conditions were optimised and the water activity was suppressed, which resulted in a yield of 38% transglycosylation products consisting of malto-oligosaccharides of various lengths. Thin layer chromatography and high-performance liquid chromatography analyses revealed the presence of malto-oligosaccharides with a higher degree of polymerisation than maltoheptaose, which has never been reported for other maltogenic amylases. The addition of organic solvents into the reaction further suppressed the water activity. The increase in the transglycosylation-to-hydrolysis ratio from 1.29 to 2.15 and the increased specificity toward maltopentaose production demonstrated the enhanced synthetic property of the enzyme. The high transglycosylation activity of maltogenic amylase offers a great advantage for synthesising malto-oligosaccharides and rare carbohydrates.

Matched MeSH terms: Oligosaccharides/biosynthesis*
Metagenomics analysis reveals significant modulation of cecal microbiota of broilers fed palm kernel expeller diets

Chen WL, Tang SGH, Jahromi MF, Candyrine SCL, Idrus Z, Abdullah N, et al.

Poult Sci, 2019 Jan 01;98(1):56-68.
PMID: 30137571 DOI: 10.3382/ps/pey366

The potential use of palm kernel expeller (PKE) as an alternative energy source in broiler diets is limited by the high fiber content. Although enzymatic treatment could alleviate the fiber component and increase the nutritive value of PKE, this apparent improvement is not reflected in the growth response of birds fed with the treated-PKE. As chicken's ceca are the most heavily populated with microflora within their gastrointestinal tract, it was hypothesized that any modulation of the intestinal environment by dietary treatments should be reflected by the composition and activities of the cecal microflora. There is a correlation between cecal microbiota composition and the efficiency of the host to extract energy from the diet and to deposit that energy into improved feed conversion ratio. At present, little is known about the changes on cecal microflora of broilers fed with PKE diets. Hence, this study was designed to assess the effects of feeding different forms of PKE; namely untreated PKE (UPKE), enzyme-treated PKE (EPKE), and oligosaccharides extracted from PKE (OligoPKE), on the cecal microbiota of broiler chickens at 14 d old (day 14) and 28 d old (day 28) using 16S rRNA gene high-throughput next-generation sequencing method. The results showed that temporal changes in cecal microbiota of broiler chickens were evident on day 14 and day 28. The relative abundance of phylum Firmicutes, known to be involved in nutrient uptake and absorption in both age groups was higher in the UPKE as compared to EPKE group. In addition, supplementation of OligoPKE increased (P < 0.05) the relative abundance of Lactobacillus on both D14 and D28, signifying its effect as prebiotics in enhancing growth of indigenous Lactobacillus. Our results showed that cecal microbiota was significantly modulated by dietary treatments and that the lower relative abundance of phylum Firmicutes in chickens fed with EPKE could be a reason why broiler chickens fed with EPKE of higher metabolizable energy (ME) content did not show improvement in their growth performance.

Matched MeSH terms: Oligosaccharides/pharmacology

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