The production of BLA can be enhanced utilizing two attacks which are the media manipulating and familial technology. Media pull stringsing involves altering the composing of the civilization media and the environing status for the agitation of bacteriums. It is of a better pick as compared to familial technology of the bacterium as the production of amylase depends on the endurance of B. licheniformis which is influenced by the civilization physical and chemical conditions ( Abdel-Fattah et al. , 2013 ) . Besides that, media manipulating is besides cheaper due to take down energy and cost input as the beginnings are readily available and easy to be obtained. Besides, media alteration allows the interaction between the parametric quantities to be studied through statistical theoretical accounts ( Sumrin et al. , 2011 ) . In contrast, familial technology can be used to do changes on the cryptography or control parts of the bacteriums amylase cistron sequence to increase its look. This method is clip and cost consuming as it involves cistron extraction, cloning of libraries, PCR, DNA assembly, sequencing and transmutation ( Leguia et al. , 2013 ) . Besides, wastage may go on due to higher opportunity of unmanageable mutants.
There are two methods for media optimisation which are the one-variable-at-a-time ( OVAT ) method and the response-surface-methodology ( RSM ) . For OVAT, merely one factor is manipulated without altering any other factors ( Soni et al. , 2012 ) . This method is less practically suited as it neglected the interactions of different factors. For blink of an eye, the bacterium may necessitate a combination of factors such as certain grade of temperature, pH and concentration of ions in order to bring forth higher sum of BLA. By altering merely one factor, this method is besides less efficient as the existent optimal status for the civilization can non be determined ( Abdel-Fattah et al. , 2013 ) . This may forestall the bacteriums from releasing maximal sum of BLA. Furthermore, OVAT is clip devouring as more experimental tests have to be performed.
In contrast, the RSM implements a set of experimental schemes, mathematics and statistical analysis which explain the interactions of different parametric quantities ( Abdel-Fattah et al. , 2013 ) . RSM is performed by transporting out experiments designed with one or more variables being manipulated and the consequences are fit into mathematical theoretical accounts to gauge the coefficients which allow the sum of BLA produced ( response ) to be estimated ( Sumrin et al. , 2011 ) . By reiterating the experiment tests, the most suited and optimal conditions to obtain higher BLA output can be determined. RSM can be carried out in different design theoretical accounts such as Plackett-Burman design ( PBD ) , Central Composite ( CCD ) and Box-Behnken design ( BBD ) where PBD is more suited for primary testing the cardinal parametric quantities while CCD and BBD are used for true finding of optimal degree for the parametric quantities identified ( Sharma & A ; Satyanarayana, 2011 ) . RSM is an alternate to OVAT as it enables the consequence of interactions between each parametric quantity to be studied. Besides, it saves clip since fewer experimental tests are needed.
The production of BLA is greatly affected by the media conditions and has to be optimized. One of the parametric quantities that have to be considered in optimising the production of BLA is the alimentary addendum such as C and nitogen. Foods are indispensable for the endurance of B. licheniformis as it determines the degree of amylase produced. This is supported by Kumar et Al. ( 2012 ) who claimed that the presence of C beginnings such as glucose, lactose and amylum or N beginnings such as ammonium nitrate and ammonium dihydrogen phosphate in civilization media will excite the BLA production. Carbon is the chief component of cellular stuffs such as cell wall and it supplies energy through glycolysis which is required for metabolic activities ( Chen et al. , 2008 ) . It is indispensable in keeping the endurance of. On the other manus, N is needed for DNA reproduction and binary fission of the bacterium as the N makes up the nucleic acid anchor of DNA and RNA. Although C and N beginnings can help in increasing BLA production, they are expensive for commercial utilizations. Hence, more economic and environmental-friendly beginnings should be used to provide foods for the bacterium to production amylase. Chandrashekhar et Al. ( 2012 ) suggested that banana wastes can be used in the agitation medium for the production of BLA as they contain high sum of sugar.
The degree of BLA produced from agitation is besides affected by the incubation temperature. Harmonizing to Mansilla et Al. ( 2004 ) , plasma membrane is made up of proteins and phospholipid bilayers with concentrated or unsaturated fatty acids. At low temperature, the hydrocarbon dress suits of fatty acids undergo trans- formation to go closely jammed ( Mrozik et al. , 2004 ) . This decreases the fluidness, doing the plasma membrane to be more stiff and there will be lesser trans-gauche isomerisation ( Mrozik et al. , 2004 ) . Consequently, the conveyance across the membrane is reduced or prevented due to decreased permeableness, taking to a lower sum of BLA produced. High temperature will blossom the 3D constructions of conveyance proteins embedded in plasma membrane, doing them to be denatured as the non-covalent interactions such as H bonds, hydrophobic and new wave der Waals interactions are broken ( Uversky, 2009 ) . The phospholipid will besides run, doing the plasma membrane to tear and take to the bacteriums cells decease. As a consequence, the degree of BLA decreased. As stated by Zare et Al. ( 2012 ) , the optimal temperature for BLA production is 37EA?C and decrease in BLA output is observed for any other temperature below or above 37EA?C. However, this consequence is opposed by Oziengbe and Onilude ( 2012 ) who claimed that the maximal the optimal temperature for B. licheniformis to turn is 60EA?C. Therefore, a more suited system should be established to happen out the most suited temperature for both the bacteriums to turn and to release more BLA.
Another parametric quantity that has to be considered in optimising the BLA production is the pH of the agitation. Sankaralingam et Al. ( 2012 ) reported that maximal production of BLA was observed at pH 7 and acidic or alkalic media will cut down the sum of BLA secreted. pH influences the growing of B. licheniformis by doing morphological alterations in the bacterium ( Kathiresan & A ; Manivannan, 2006 ) . Besides, it will impact the solubility of foods which is required for bacterial growing, ions bindings and metabolic activities such as synthesis of ATP which determine the endurance of bacteriums ( Slonczewski et al. , 2009 ) . Besides, pH will impact the amylase stableness in the media. Amino acids are zwitterions and they are connected through non-covalent interactions such as H bonds, disulphide Bridgess, hydrophobic interaction and new wave der Waals interaction to organize a 3D construction of BLA. When there is a lessening in pH, the concentration of H ions is higher and these H ions will adhere to the carboxyl group of the amino acid, doing conformational alterations in BLA as the interactions are disrupted. This appears to be the same for alkalic status where the hydrated oxide ions will do the amino group to lose a H ion. Consequently, the BLA is denatured taking to a lower sum of output obtained.
In decision, it is really of import to optimise the agitation of B. licheniformis in order to obtain maximal output of BLA. Media use is better in optimising BLA production as it is cheaper and simpler compared to familial technology which is more expensive due to complex processs and instrumentality and it is clip devouring. Media optimisation can be carried out utilizing OVAT and RSM methods but RSM is more preferable as it allows the true optimal status to be determined from the interactions of different parametric quantities. In order to increase the sum of BLA produced, handiness of C and N beginnings, incubation temperature and the pH of the media have to be considered as these parametric quantities influence the endurance of bacteriums and concluding sum of BLA. However, future surveies have to be carried out to farther look into the optimal conditions for BLA production as there are still restrictions on present methods.