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What he doesn't know is what taking on another case might cost him this time. Table of Contents The Gifted Realm: Chapter 36 - It All Comes Tumbling Chapter 65 - Dan vs.

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New to the Gifted Realm? Check out the handy Reference Guide Want to go back to where it all began? Tip Cup I hope you are enjoying reading. Feel free to throw a little bit in the tip cup to keep me caffeinated and cranking out new chapters. Just click the coffee mug to donate. TA lipases production was also improved through recombinant techniques. The TA lipase gene from T. Another lipase gene like Gene lipZ01 was expressed in P. On the other hand, lipase gene from Cohnella sp. Lip 42 gene from Bacillus sp. Some recombinant proteins produced by E. When it comes to downstream processing, extracellular lipases may be preferred as compared to intracellular lipases due to the tedious cell disruption process in order to release the enzymes.

In response of surface methodology RSM optimization study, a very low inoculums size was suggested for the production of lipase from Acinetobacter sp. Meanwhile, the production of TA lipase from B. It has been reported that Bacillus sp. LBN2 at inoculums size of 10 4 cells per mL was inoculated into medium for maximal lipase activity [ 9 ]. It is essential to determine the optimum inoculums size for optimal number of active microbial cells needed for TA lipases production.

Large inoculums size not only can cause overproduction of microbial mass and inefficient mass transfer but also is unsuitable for scale up process and production in large bioreactor. A variety of carbon and nitrogen containing substrates could be used as carbon and nitrogen sources for TA lipases fermentation. The type, amount, and ratio of carbon and nitrogen sources are important for microbial growth and optimal production of TA lipases.

The most favourable carbon sources for TA lipase production by various thermophilic microbes are varied. Olive oil containing large percentage of monounsaturated fats was mostly used as carbon source and in some cases as an inducer for lipase production. For instance, TA lipase was optimally produced by B. This is potentially due to TA lipase substrate specificity to unsaturated fatty acid. In addition to this, isolate B. Another lipase production by Bacillus species like B. TA lipase was also produced by anaerobic T.

TA lipase production was optimally produced by G. Organic nitrogen source like yeast extract improved not only TA lipase production by T. The production of TA lipases from T. This has been supported by several works on reduction of lipase production by many sugars and complex substrates at higher concentrations [ 26 , 34 ]. Peptone was found to be the most suitable nitrogen source for TA lipase from G.

Olive oil is the best source of oil which can act as an inducer for the production of lipase from Bacillus stearothermophilus , followed by nut oil, sesame oil, sunflower oil, Tween, Triton-X, and Tween [ 26 ]. High TA lipase from B. On the other hand, high TA lipase yield by B. Pumilus HF can be produced using a mixture of peptone and yeast extract as nitrogen sources and glucose as carbon source [ 33 , 60 ].

Moreover, lipases activity of some other fungi, such as B. Although TA lipase production from this strain is constitutive, the incorporation of oils in the medium increased its final yield. The yield was greater in a medium containing lipidic substrates such as oils as the carbon sources with an addition of organic nitrogen source, but, sometimes, lipase production was also repressed by polysaturated, long chain fatty acid LCFA and esters [ 17 , 46 , 74 ].

RSM optimization for lipase production by Acinetobacter sp. AU07, suggests a significant increase in lipase yield using castor oil as an inducer 2. The chain length specificity of the TA lipase gene from T. Lipases show very low activity when substrates are in the monomeric form as lipases are activated at the water-lipid oil interface.

The activity increases significantly when substrates form emulsions due to interfacial activation. This explains the necessity of emulsions for maximum lipolytic activity for the majority of lipases [ 42 , 83 ]. Additional minerals were required for TA lipase stability and to boost TA lipases production. TA lipase production by B. It was found that lipase stability of B.

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Certain metal ions give positive effect to TA lipases synthesis and stability due to the presence of metal binding motif in its structure. Some other minerals such as phosphate ions and NaCl provide ATP synthesis and osmotic balance for cell growth and TA lipases production.

The optimum growth condition i. Temperature regulates and gives an effect to the lipase synthesis at mRNA transcriptional regulation of lipase gene and also translation levels of lipase proteins. This involves several other proteins along the way such as regulatory protein, polymerase i. The optimum growth condition and lipase production by T. Under optimized condition, high lipase production by Acinetobacter sp.

Higher temperature for microbes mentioned above was not however favourable for TA lipases production, probably due to denaturation of regulatory peptides and proteins, responsible for lipases production.

• Realm of Thermoalkaline Lipases in Bioprocess Commodities.
• INTRODUCTION;
• Naughty: Twos Enough, Threes a Crowd.

High temperature may also disturb the stability of cell membranes structure as not all microbes have a heat-tolerant cell membrane [ 41 ]. Studies show that optimal temperature for production of TA lipase may not correlate to the temperature of the TA lipases activity, which is normally active and stable at a higher temperature. Other TA lipases are highly produced at high temperature and this includes TA lipase production by B.

A very high TA lipase production by B.

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High TA lipase was significantly produced by T. Most of the microbes are thermopiles or hyperthermophiles and capable of withstanding and growing at a very high temperature. High TA lipases at these temperatures are usually related to high kinetic rate and mass transfer rate MTR at high temperature [ 82 ]. An elevated temperature may influence their secretion, possibly by changing the physical properties of the cell membrane, thus allowing more secretion of extracellular lipases [ 56 ]. In shake flask culture SFC , agitation rates between and rotations per min rpm have been reported for the production of TA lipases.

For instance, TA lipase production by T. The production of TA lipase Acinetobacter sp. The lipase production by B. Due to geometry difference of shake flask and bioreactor, the effect of agitation rate on lipases production was normally reevaluated in bioreactor. Furthermore, in bioreactor, such as stirred tank bioreactor STB , agitation rate is controlled by one or more impellers. The effect of different impeller design may give some other effect such as shear stress especially at a very high agitation rate which may damage cells and pulpy state moulds [ 41 ].

Medium pH is very significant in nutrients absorption and growth of microorganisms, stimulation of lipase production via signaling pathways, and release of extracellular lipases. In SFC, the effect of pH on the production of TA lipases was solely conducted at initial pH of the fermentation medium and pH was not controlled throughout the fermentation time due to small fermentation size. The initial pH of 6.

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TA lipase production was also high at initial alkaline fermentation pH by T. The lipase production of Bacillus sp. LBN2 was found high at alkaline pH range of 8. It was suggested by RSM that a significant increase in lipase production by Acinetobacter sp. AU07 could be achieved at fermentation pH 7. The optimum initial pH for maximal production of TA lipases by various microbes is varied maybe due to their nature and habitat where they were isolated.

Initial pH of the fermentation medium serves as starting pH for the microbes to grow. Initial pH was usually enough for high production of TA lipases and addition of acid or base to control pH medium at certain value may inhibit growth and production of TA lipases. The microbes will slowly adapt to the changes in fermentation medium as pH drop or rise which explains the reason that medium was only set to an initial pH. In few cases, some TA lipases production can be optimized at controlled pH of the fermentation medium [ 41 ].

Aeration rate and dissolved oxygen tension DOT are important parameters as most microbes reported in literatures were aerobes or facultative anaerobes. The influence of aeration rate cannot be studied in SFC but it can be controlled in bioreactor i. In SFC, aeration rate and dissolved oxygen DO are solely determined by agitation rate but this is not the case for bioreactor system where the agitation and aeration rate can be set separately and maintained at a desired value [ 92 ].

However, in bioreactor system, DOT can be monitored using DO probe and controlled throughout the fermentation [ 93 ]. In bioreactor system, the production of TA lipases was enhanced at aeration rate of 1. Fermentation of Acinetobacter sp. These reported studies have demonstrated that the effect of aeration rate and DOT in bioreactor play a major role and responsible for the increase in growth of the aerobes and production of the TA lipases.

During growth phase and in a situation where oxygen MTR was limited, moulds cells in the form of mycelia pellet may be prone to autolysis and caused void formation in the interior part of the pellet [ 41 ]. The TA lipases production in the nonagitated fermentation culture can also be restricted by MTR where oxygen uptake by the cells was scarce [ 92 , 93 ]. TA lipases production in SFC has its own limitation due to the fact of its small size and geometrically not scalable.

A successful optimization in small scale fermentation condition was needed for further study in pilot and industrial scales for possible industrial-scale production via bioreactor system Table 6. Conventional and new bioreactors as well as impeller designs were studied and compared in terms of their efficiency and feasibility for large-scale TA lipases production. TA lipase production by an anaerobic T. On the other hand, submerged fermentation SmF of Acinetobacter sp.

In SFC, the recombinant E.

Moreover, lipase production by S. Under these conditions, the cell concentration reached its maximal value of 8. For instance, lipase production from fungus strain i. In industry, cost and simplicity of the production techniques are crucial factors for mass-scales TA production. The feasibility of using various types of fermentation techniques i. Generally, SSF is known as a method where fungus and moulds are grown on a solid medium with limited free liquid phase of the culture [ 41 , 97 ].

For example, a comparatively high activity of Aspergillus terreus lipases MW of Meanwhile, the thermotolerant Rhizopus homothallicus produced more lipases MW of The lipase production In contrast to SmF, the SSF process occurs in very low water content and thus the lipases are strongly adsorbed on the insoluble biomass, which later require an efficient extraction process and lipase recovery.