TECHNOLOGY FOCUS - BIOTECHNOLOGY - Preparation: (I) which exhibits a reduced capability of cleaving a substrate close to the branching point, and further exhibits improved substrate specificity and/or improved specific activity relative to the parent, is generated by a process which involves subjecting a DNA sequence encoding the parent TLA to random mutagenesis, expressing the mutated DNA sequence in a host cell and screening for host cells expressing a mutated alpha-amylase which has increased stability at low pH and calcium concentration relative to the parent alpha-amylase (claimed). Preferred Variant: (I) comprises a mutation in a position, corresponding to one of the following mutations in (S4) such as V54N, A52S, A52S+V54N, T49L, T49L+G107A, A52S+V54N+T49L+G107A, A52S+V54N+T49L, G107A, Q51R, Q51R+A52S, A52N; or T49F+G107A, T49V+G107A, T49D+G107A, T49Y+G107A, T49S+G107A, T49N+G107A, T49I+G107A, T49L+A52S+G107A, T49L+A52T+G107A, T49L+A52F+G107A, T49L+A52L+G107A, T49L+A52I+G107A, T49L+A52V+G107A; or T49V, T49I, T49D, T49N, T49S, T49Y, T49F, T49W, T49M, T49E, T49Q, T49K, T49R, A52T, A52L, A52I, A52V, A52M, A52F, A52Y, A52W, V54M, G107V, G07I, G107L, G107C. (I) also comprises a mutation in a position corresponding to one of the following mutations in the amino acid sequence of (S4). W13F, L, I, V, Y, A; G48A, V, S, T, I, L; asterisk 48aD or 48aY; T49X; asterisk49aX (i.e. insertion of any amino acid residue) S50X, in particular D,Y,L,T,V,I; Q51R, K; A52X, in particular A52S, N, T, F, L, I, V; D53E, Q, Y, I, N, S, T, V, L; V54X in particular V54I, N, W, Y, F, L; G57S, A, V, L, I, F, Y, T; G107X, in particular G107A, V, S, T, I, L, C; G108X, in particular G108A, V, S, T, I, L; A111V, I, L; S168Y; M197X in particular Y, F, L, I, T, A, G. Also it comprises a mutation of T49X+A52X+V54N/I/L/Y/F/W+G107A. (I) also comprises the following mutations. V49L+G107A; V49I+G107A; V49L+G107A+V54I; V49I+G107A+V54I; A52S+V54N+T49L+G107A; A52S+V54I+T49L+G107A; A52S+V54I+T49L+G107A; A52S+T49L+G107A; A52T+T49L+G107A; A52S+V54N+T49I+G107A; A52S+V54I+T49I+G107A; A52S+T49I+G107A; T49L+G107A; T49I+G108A; T49L+G108A+V54I; T49I+G108A+V54I. The parent hybrid alpha-amylase is a hybrid alpha-amylase comprising the 445 C-terminal amino acid residues of the B. licheniformis alpha-amylase having a sequence of (S4) and the 37 N-terminal amino acid residues of the alpha-amylase derived from B. amyloliquefaciens having a sequence of (S6). The parent TLP alpha-amylase as the following mutations: H156Y+A181T+N190F+A209V+Q264S or LE174. Alternately, the TLP comprises the mutations H156Y+A181T+N190F+A209V+Q264S+I201F or LE429.Preferred Host Cell: (IV) is a microorganism such as a bacterium or fungus, such as gram positive bacterium e.g. B. subtilis, B. licheniformis, B. lentus, B. brevis, B. stearothermophilus, B. alkalophilus, B. amyloliquefaciens, B. coagulans, B. circulans, B. lautus or B. thuringiensis.Preferred Composition: In (VIII) the hybrid alpha-amylase comprises the 445 C-terminal amino acid residues of the B. licheniformis alpha-amylase having a sequence of (S4) and the 37 N-terminal amino acid residues of the alpha-amylase derived from B. amyloliquefaciens having a sequence of (S6). The parent TLP alpha-amylase as the following mutations: H156Y+A181T+N190F+A209V+Q264S or LE174. Alternately, the TLP comprises the mutations H156Y+A181T+N190F+A209V+Q264S+I201F or LE429. EXAMPLE - Termamyl Bacillus licheniformis alpha-amylase having a fully defined sequence of 483 amino acids as given in specification was expressed in B. subtilis from a plasmid denoted pDN1528. A specific mutagenesis vector pJeENI containing a major part of the coding region which has a fully defined sequence of 1920 nucleotides as given in the specification was prepared. Mutations were introduced in amyL by the method described by Deng and Nickoloff (1992, Anal. Biochem. 200, pp. 81-88) except that plasmids with the selection primer incorporated were selected based on the ampR phenotype of transformed Escherichia coli cells harboring a plasmid with a repaired bla gene, instead of employing the selection by restriction enzyme digestion. After verification of the DNA sequence in variant plasmids, the truncated gene, containing the desired alteration was subcloned into pDN1528 as a PstI-EcoRI fragment and transformed into the protease- and amylase-depleted B. subtilis strain SHA273. The termamyl variant V54W was constructed by the use of the following mutagenesis primer: PG GTC GTA GGC ACC GTA GCC CCA ATC CGC TTG. The termamyl variant A52W + V54W was constructed by the use of the following mutagenesis primer: PG GTC GTA GGC ACC GTA GCC CCA CCA TTG GCT CG. Prime 6616. P CTG TGA CTG GTG AGT ACT CAA CCA AGT C. The termamyl variant V54E was constructed by the use of the following mutagenesis primer: PGG TCG TAG GCA CCG TAG CCC TCA TCC GCT TG. The other termamyl variants are produced by employing a similar procedure.