The use of pullulanase (EC 3.2.1.41) has recently been the subject of increased applications in starch-based industries especially those aimed for glucose production. Pullulanase, an important debranching enzyme, has been widely utilised to hydrolyse the α-1,6 glucosidic linkages in starch, amylopectin, pullulan, and related oligosaccharides, which enables a complete and efficient conversion of the branched polysaccharides into small fermentable sugars during saccharification process. The industrial manufacturing of glucose involves two successive enzymatic steps: liquefaction, carried out after gelatinisation by the action of α-amylase; saccharification, which results in further transformation of maltodextrins into glucose. During saccharification process, pullulanase has been used to increase the final glucose concentration with reduced amount of glucoamylase. Therefore, the reversion reaction that involves resynthesis of saccharides from glucose molecules is prevented. To date, five groups of pullulanase enzymes have been reported, that is, (i) pullulanase type I, (ii) amylopullulanase, (iii) neopullulanase, (iv) isopullulanase, and (v) pullulan hydrolase type III. The current paper extensively reviews each category of pullulanase, properties of pullulanase, merits of applying pullulanase during starch bioprocessing, current genetic engineering works related to pullulanase genes, and possible industrial applications of pullulanase.

Lactic acid bacteria (LAB) are of major interest in the food industry primarily by virtue of their biopreservative properties. LAB have ability to produce various types of antimicrobial compounds, the most important being bacteriocins. Bacteriocins and bacteriocin-producing cultures have the potential to increase the shelf-life of foods and contribute towards decreasing the incidence of food-borne diseases. In this respect, food preservation through in situ production of bacteriocins by LAB introduced into the food system would be the most logical approach. However, there is a need to understand the relationship between bacterial growth and bacteriocin production in various types of food system. Bacteriocin production by LAB is dependent on a number of factors such as the types of carbon and nitrogen sources and their concentrations in the media formulation. Other factors which need to be considered are the culture conditions which include pH, temperature and aeration which greatly influence the cultivation performance of bacteriocins producing LAB. Economic aspects pertaining to the optimization of fermentation process for the enhancement of bacteriocin production should also be given due considerations. Failure to acknowledge or recognize this hidden economic element would be a substantial financial loss to the industry especially from the point of view that the product is costly and highly sought after. Thus, the fermentation factors which influence the production of bacteriocins by LAB and the approaches to improve the production not only in term of yield and productivity but also in term of economic and regulation are reviewed in this paper.

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BackgroundLactic acid bacteria (LAB) can be isolated from traditional milk products. LAB that secrete substances that inhibit pathogenic bacteria and are resistant to acid, bile, and pepsin but not vancomycin may have potential in food applications.ResultsLAB isolated from a range of traditional fermented products were screened for the production of bacteriocin-like inhibitory substances. A total of 222 LAB strains were isolated from fermented milk products in the form of fresh curds, dried curds, and ghara (a traditional flavor enhancer prepared from whey), and fermented cocoa bean. Eleven LAB isolates that produced antimicrobial substances were identified as Lactococcus lactis, Lactobacillus plantarum, and Pediococcus acidilactici strains by biochemical methods and 16S rDNA gene sequencing. Of these, the cell-free supernatant of Kp10 (P. acidilactici) most strongly inhibited Listeria monocytogenes. Further analysis identified the antimicrobial substance produced by Kp10 as proteinaceous in nature and active over a wide pH range. Kp10 (P. acidilactici) was found to be catalase-negative, able to produce β-galactosidase, resistant to bile salts (0.3%) and acidic conditions (pH 3), and susceptible to most antibiotics.ConclusionTraditionally prepared fermented milk products are good sources of LAB with characteristics suitable for industrial applications. The isolate Kp10 (P. acidilactici) shows potential for the production of probiotic and functional foods.

Optimization strategy that encompassed one-factor-at-a-time (OFAT), response surface methodology (RSM), and artificial neural network method was implemented during medium formulation with specific aim for lysine-methionine biosynthesis employing a newly isolated strain of Pediococcus pentosaceus RF-1. OFAT technique was used in the preliminary screening of factors (molasses, nitrogen sources, fish meal, glutamic acid and initial medium pH) before proceeded to optimization study. Implementation of central composite design of experiment subsequently generated 30 experimental runs based on four factors (molasses, fish meal, glutamic acid, and initial medium pH). From RSM analysis, a quadratic polynomial model can be devoted to describing the relationship between various medium components and responses. It also suggested that using molasses (9.86 g/L), fish meal (10.06 g/L), glutamic acid (0.91 g/L), and initial medium pH (5.30) would enhance the biosynthesis of lysine (15.77 g/L) and methionine (4.21 g/L). Alternatively, a three-layer neural network topography at 4-5-2 predicted a further improvement in the biosynthesis of lysine (16.52 g/L) and methionine (4.53 g/L) by using formulation composed of molasses (10.02 g/L), fish meal (18.00 g/L), and glutamic acid (1.17 g/L) with initial medium pH (4.26), respectively.