Jor band migrating at 49 kDa, plus some minor bands of decrease mass.18 The meals enzyme was tested for protease, b-glucanase, phospholipase and glucoamylase activities; none had been detected.19 The in-house determination of a-amylase activity is determined by hydrolysis of 4,6-ethylidene(G7)-pnitrophenyl(G1)-a-D-maltoheptaoside (ethylidene-G7pNP) by a coupled reaction that results in the release of p-nitrophenol (reaction conditions: pH 7.0, temperature 37 , reaction time five min). The enzymatic activity is quantified by measuring the formation of p-nitrophenol spectrophotometrically at 405 nm. The activity is quantified relative to an internal enzyme standard (S) and expressed in Kilo Novo alpha-amylase Units (S)/g (KNU(S)/g).20 The meals enzyme includes a temperature optimum around 70 (pH 4.five) and also a pH optimum about pH 5 (30 ). Its thermostability was tested just after a pre-incubation for 30 min at diverse temperatures (pH four.5). The a-amylase activity decreased above 70 , displaying no residual activity at 100 .three.three.two.Chemical parametersData on the chemical parameters from the meals enzyme have been supplied for 3 batches used for commercialisation and a single batch made use of for toxicological testing (Table 1). The mean total organic solids (TOS) of the three meals enzyme batches made use of for commercialisation was four.0 plus the imply enzyme activity/TOS ratio 24.six KNU(S)/mg TOS.5 Table 1: Compositional information of the food enzymeBatch Parameters a-amylase activity Protein Ash Water Total organic solids (TOS)(c) Activity/mg TOS Unit KNU(S)/g batch(b) KNU(S)/mg TOS 1 1,220 three.0 0.6 94.7 4.7 26.0 two 632 2.0 0.4 97.0 2.6 24.three 3 1,130 three.three 0.four 94.eight four.8 23.5 4(a) 1,760 7.two 3.1 87.0 9.9 17.(a): Batch employed for the genotoxicity studies. (b): KNU(S): Kilo Novo alpha-amylase Units (see Section 3.three.1). (c): TOS calculated as one hundred water ash.3.three.3.PurityThe lead content inside the 3 commercial batches and within the batch employed for toxicological research was below 0.5 mg/kg, which complies together with the specification for lead ( five mg/kg) as laid down inside the common specifications for enzymes employed in meals processing (FAO/WHO, 2006).Orexin A Neuronal Signaling 5 The levels of cadmium, mercury and arsenic were under the limits of detection (LODs) with the employed methodologies.Atipamezole Adrenergic Receptor 22,23 The meals enzyme complies with all the microbiological criteria (for total coliforms, Escherichia coli and Salmonella) as laid down in the basic specifications for enzymes utilized in food processing (FAO/WHO, 2006).PMID:23892746 24 No antimicrobial activity was detected in any from the tested batches (FAO/WHO, 2006).25 The Panel regarded that the data provided around the purity from the meals enzyme was enough.18 19 20 21 22 23 24Technical dossier/page 60. Technical dossier/pp. 68. Technical dossier/pp. 635/Annex three. Technical dossier/page 667/Annex 9. LoDs: Pb = 0.5 mg/kg; As = 0.1 mg/kg; Cd = 0.05 mg/kg; Hg = 0.03 mg/kg. Technical dossier/pp. 134 and 61. Technical dossier/pp. 15 and 63. Technical dossier/pp. 14 and 61. eight EFSA Journal 2022;20(7):efsa.europa.eu/efsajournalSafety with the a-amylase in the genetically modified Bacillus licheniformis strain NZYM-BC3.three.four.Viable cells and DNA with the production strainThe absence of viable cells with the production strain in the food enzyme was demonstratedThe absence of recombinant DNA within the meals enzyme was demonstrated3.four.Toxicological dataThe applicant supplied a bacterial gene mutation assay (Ames test) and an in vitro micronucleus test performed with the food enzyme beneath assessment (batch 4, Table 1). Additionally, a.
