Revolutionary Era Paper Essay Example | Topics and Well Written Essays - 750 words

Revolutionary Era Paper - Essay Example Thirteen colonies in North America joined and broke free from the British Empire, merged, then became the United States of America. They deterred the rules and regulations of Great Britain Parliament that governed them from overseas with no representation. A number of ideas sparked this revolutionary era. During this period, the idea of liberty fueled Americans into actions that sparked revolution in their country2. What the eighteenth-century politicians and other people meant by liberty was, first, a wake up call for individual human rights. This idea was so radical because even the most enlightened and revised monarchs believed customarily that it was their task to keep an eye on what people wrote or believed. Liberals of this revolutionary era protested those particular controls that enslaved their conscious. According to Americans, liberty was of overriding importance to them since they believed it would be through liberty, they would get to choose and define the outcome of thei r censorship. They demanded freedom from arbitrary laws and judges who commonly and easily obeyed rules from the government that did not carry the rule of law to the letter. Their feelings towards freedom of worship with regard to their consciences acted as a morale booster towards revolution. This eventually led to freedom of worship amongst other things. ... However, many eighteenth century men were actually individuals of some kind. They generally shared with other men the belief that it was neither practical nor desirable for women to be their equals. Therefore, they curtailed their political rights. They could not vie for office or participate in the governing of the state. Again, liberals despised the belief that everyone should be equal economically. They based this on the belief that, not everyone could succeed equally in pursuit of acquiring property during American conquest for independence. Hence, great rift in terms of wealth or material possession and income generation between the rich and the poor was highly encouraged and perfectly acceptable by liberals. British administrators taxed Virginia citizens. In 1765, after North America restored peace, the British government decided to push via parliament the Stamp Act that sought to levy taxes on a longer list of legal documents and commercial articles like pamphlets, diplomas, a nd newspapers. Effort to end taxation started as Americans claimed taxation without representation was tyranny. This idea rose questions as to who should make laws for American people. Colonial experience formed people who felt that they were separate and distinct to their very own country. The taxation controversies made these feelings separation and distinctiveness more intense thus bringing them to the fore. These extreme attempts led to expanded governor’s power. As fighting spread and the colonialists moved slowly, East Virginia inhabitants developed ideas of being independent. They inevitably opened up towards open rebellion and boldly shown urge to declare their independence. Some American icons arranged civilized and

Employee Turnover and Retention Strategies Essay

Employee Turnover and Retention Strategies - Essay Example Every department, be it operations, finance, human resources, or sales and marketing, must invest in motivating, training, and retaining their best human resources to make a difference through their core competencies. As Susan Heathfield said: "Managers readily agree that retaining your best employees ensures customer satisfaction, product sales, satisfied co-workers and reporting staff, effective succession planning and deeply imbedded organizational knowledge and learning." Today organisations are willing to look upon their staff not so much as employees or workers but as 'knowledge workers'. As Peter Drucker observed, "while the more than fifty-fold increase in productivity of the manual worker was the greatest achievement of the 20th century manager, the most important work of the 21st century is to increase the productivity of the knowledge worker." (Pottruck and Pearce, 131). The more competent workers your company or organisation loses, the more will it set your clock back in finding and training suitable replacements at much greater costs. In the meantime your loss becomes your competitor's gain, which again has adverse implications on the success of your operations. Among the major causes that propelled employees to quit an organisation, the following conditions mostly relating to work culture have been identified as contributing to employee dissatisfaction and consequent exit: Lack of clarity in company policies, practices, and procedures Excessive controls and autocratic leadership Bureaucratic work environment with lack of trust Inadequate remuneration and unjustified cost cuts Poor quality of supervision Lack of opportunities for growth Absence of facilities for training and development Lack of scope for utilization of skills and talents Poor communication (with the employee not knowing what is expected of him daily) Lack of rewards, recognition and encouragement Unfair and inequitable treatment of employees Overbearing attitude and threatening by supervisors The Case of Solution's Solution's began operations on a small scale in 1967 as a recruitment agency for social care specialists. Headquartered in London, Solution's expanded in a big way in four decades to establish itself as the largest social care agency in the UK, with 160 employees spread over18 branches. The year 2004 proved to be a disappointment for the company in terms of financial performance, compelling it to slash commissions given to consultants for recruitment mostly in branches located in London. The weak performance also resulted in the retrenchment of up to 15 branch administers, apart from the voluntary exit of many employees who signed up with rival agencies. An analysis showed that the major cause for high employee turnover (91%) in 2004 was the cost reduction strategy adopted by the company. With a view to putting the operations back on rail, the company opted for a change of leadership at the top. A new managing director joined the board in January 2005 with a mission to improve the health of the company in general and the turnover situation in particular. He believed that the key to success through increased sales lay in ensuring employee satisfaction. Accordingly he straightway set about implementing certain remedial and innovative measures for a

Toni Morrison Is Adulthood inevitable Essay Example | Topics and Well Written Essays - 1250 words

Toni Morrison Is Adulthood inevitable - Essay Example She stated that each phase of individual’s life is very important and has to be experienced with the greatest enthusiasm. Since she herself is an adult now, and she practices what she preaches, so there is nothing more satisfactory than becoming the adult in mind and heart. Morrison explained brightly that adulthood is about being content with the present, happy with the rewards one gets, and also work hard every day to make ones adulthood fulfilling. I completely agree that an individual needs to be in the present moment and be happy in that because the present is what matters the most since the past is gone and future is yet to be experienced. Living this way not only enlightens the present, but also the past and the future since one has lived every moment happily as the past and such a philosophy lays the basis of a bright future ahead as well. Morrison defined true adulthood in terms of the ability of an individual to take responsibility for his/her actions. This only can be achieved if people understand their role in the society in addition to just thinking about their personal well-being. It is not the chronological age, white hair or time period that defines adulthood. Adulthood is defined by an individuals attitude and approach towards life. As individuals, we have to choices. First; we can be selfish and take care of our own needs, and safeguard only our own interests, and second; we can live for more than ourselves, that is to say that we live for others and try to make a positive change in their life. If we adopt the first approach towards life, adulthood for us does not get inevitable. But if we choose to adopt the second approach towards life, adulthood for us gets inevitable. Hence, to say that adulthood is inevitable and generalize it for all people is wrong because there are many people who live and care only for themselves, so they are not included in this category. However, for the rest, who live and care for others, it can

Apple, Samsung ,Nokia and blackberry smart phones comparison between Essay

Apple, Samsung ,Nokia and blackberry smart phones comparison between these 4 companies - Essay Example This has further led to the fast embracing of modern technology handheld gadgets like the Smart phones and tablets and I pads, which provide people all over the world with the virtual internet based platform to connect with their friends and relatives located in various parts and corners of the world. The changing lifestyle trends of the masses all over the world along with the desire to stay connected and aware of the happenings and events in the lives of their near and dear ones by the process of getting updates in their social networking profiles has also played a major role in the tremendous acceptance of Smart phones all over the world. As a matter of fact, in an order to capitalize on the growing demands of data based technology services like the internet in the recent times, various telecom service providing companies has focused on the process of bundled offering of products and services, thereby offering the customers with high value internet services for their mobiles and gadgets at very competitive rates. It can be said that the global smart phone market is currently dominated by two of the largest and most popular mobile manufacturers namely Apple Inc. and Samsung Electronics, while talking in terms of revenue generation, sales and share of the markets in the global arena. Talking from the statistical point of view, it can be said that the shipping statistics of the second quarter of the year 2012 highlighted that Apple iOS and Samsung Android based smart phones comprised almost 85% of the total shipments (IDC.com -1, 2012)1. Talking about the market sentiments that was existing around the middle of 2012, during the period of launch of I-phone 5 in the United States, a research which was undertaken by AYTM along with Mashable stated that current owners of Android phones in the United States are looking forward to move to I-phones within the next 6 months (Price, 2012)3 . The survey also revealed that

International relations Article Example | Topics and Well Written Essays - 1250 words

International relations - Article Example This section uses realism theory to explain relationships among state and non-state actors in the international system. Realism is based on the idea that the international system exists according to the Hobbestian state of nature, which is anarchic and brutal. There is no centralized authority in the system. The influential states have powers that they use to subjugate the poor and powerless in the system. The paper evaluates three supranational regimes in order to understand the theory in detail. States agree to form governments that control internal affairs. It is not surprising that any sovereign state has the power to enforce and prescribe laws. These powers are limited in the international system because there is no Leviathan that controls the activities of all members. The manner in which African states and other developing countries conduct politics is a matter of great concern when it comes to the issue of classical realism. From liberalists’ point of view, the drives to power and will to dominate is based on liberty in which peoples’ freedom to determine the nature of politics is given priority in the society. According to Kenneth Waltz and Wendt, people are free to elect the leader they want. They are also at liberty to contribute to the government without restrictions. Neorealist focuses on the international system in which states are considered as the key actors. ... The difference between classical realism and neo-realism is based on how they view the causes of conflict in international relations. Classical realism view unchanging human nature and self-interest as the major causes of conflict in international relations while neo-realism consider state anarchy as the major cause of conflict in international relations (Harrison 89). Question 3: Persistent underdevelopment The dependency theory is utilized in international relations to predict dependency in third world countries. Dependency has been defined as a condition in which the growth and expansion of an economy is conditioned or determined by growth and expansion of another economy. This means that a dependent state cannot experience a self-sustaining development that is; it cannot be self-reliant because its economy is a reflection of the developed state’s economy. The process of dependency is sustained by foreign aid including technical assistance and military aid. The aid does not enhance the economy but it strengthens dependency relations. The aid further marginalizes the dependent countries since they create permanent debts. Marxists argue that neither mercantilism nor liberalism guarantee human happiness this is because the owners of the means of production control the markets whereas the state is the property of the elite. It therefore postulates that people should determine their own destiny through collectivization of interests (Allen and Thomas 69). Liberalism argues that politics and economics have no relationship whatsoever. The market according to liberalists is self-regulating because it operates on its internal logics. It claims that the market

Setting Up A Moral System Essay Example for Free

Setting Up A Moral System Essay The installation of a moral system is vital in every society. Yet, every moral system must deal with the major conflicting general moral issues: Consequentialism versus Non-consequentialism; Self versus Other-Interestedness; Act Utilitarianism versus Rule Utilitarianism; and Emotion versus reason. When all four areas are combined and examined there are conflicting views that must be resolved. First of all, there is the issue of Consequentialism and Non-consequentialism. Consequentialism is best described by utilitarianism, where decisions are based on the consequences of the outcome of the decision made. On the other hand, non-consequentialism is best described as an ethical theory that is based not on the consequences of the action, but on some higher moral standard that is placed upon them. Examples of non-consequentialism are Kants Duty Ethics and Divine Command Theory. The conflict between the two is whether to do what is best according to the outcome or follow what one feels as the higher moral standard would require them to adhere to. Personally, I would resolve this area by choosing the consequentialist belief as my tool for judgement because I feel that people need to think about the consequences of their actions because if they dont then it will cause havoc in the society. Thus, I feel that consequentialism is the proper solution in this area. Secondly, there is the conflict between Self versus Other-Interestedness. The concept of self is best described as being selfish and only caring about oneself without worrying about what might happen to others. While the concept of other-interestedness is basically putting the needs of everyone else over your own (this is similar to utilitarianism). The big conflict is that most people are going to choose themselves first, but if this were a perfect society the people would worry about everyone first. If people would think about it, then they figure it out that they are included in everyone. Therefore, in this area I would solve the problem by choosing the theory of other-interestedness. Next, there is the issue of the act utilitarianism versus the rule utilitarianism. The act approach to utilitarianism is that a person should perform acts that will bring about the greatest good for the greatest number. On the other hand, the rule approach to utilitarianism is that people establish and follow rules which will bring about the greatest good for the greatest number. The major conflicting moral issue is whether or not there is freedom involved in the decision making process. Act utilitarianism gives unlimited freedom, while rule utilitarianism gives virtually no freedom, but rule utilitarianism gives structure and stability to the society. The best way to solve this area would be to have rule utilitarianism and act utilitarianism combined so that you have rules to give some structure to society but have the ability for the people to be allowed to express their individual freedoms in the society. Thus, I feel that a combination of the two is the best way solve this area of the moral system. Finally, there is the issue between reason and emotion. Reason is described as thinking about what the outcome of the decision would be, then making the decision. The theory of emotion is reacting to ones emotions without thinking about the possible outcome of the situation. In this situation one has to side with reason because if not the society would have people reacting to their emotions all the time which could chaos and would not fit into a utopian society. All of these four areas combined would help in establish an upright and proper moral system when applied correctly. I feel that the decisions that I made earlier in this essay are the proper ones needed for a good moral system.

Production of Pure Lipases

Production of Pure Lipases Most Rhizopus lipases described in the literature are produced by their host cells in very small amounts, which is not suitable for large-scale industrial applications. Another drawback of working with native microorganisms is that Rhizopus sp. usually produces different isolipases with different biocatalytic properties, which complicates biocatalytic application with unwanted side effects caused by unrelated enzymes (67). Thus, in order to implement more industrial lipase applications, one of the most promising strategies is to produce heterologous lipases in a host microorganism which have the advantages to minimize the complexity of the bioprocess, to facilitate the more economic production of large quantities of pure lipases and to reduce costs (68). Rhizopus species was mainly divided into three groups, including R. oryzae, R. microsporus, R. stolonifer (69). Until now, the lipase genes from all these three groups have been cloned. Consistent with the proposal that many Rhizopus isolates are actually the same organism (69), nearly complete identities were observed in the lipase genes from the genus Rhizopus. In the R. oryzae group, lipases from R. niveus IFO 9759 (70) (71) (72), R. delemar ATCC34612 (73) (74) and R. javanicus (75) have identical amino acid sequences, and the lipases from R. oryzae ATCC 853 (76, 77) (78) and R. arrhizus L-03-R-1(79) differ by only several substitutions. The lipase from R. stolonifer (GenBank No. AAZ66864.1) and R. chinensis (GenBank No. ABN59381.2)(80) were of about 80% amino acid sequence identity to R. oryzae lipase (GenBank No. BAG16821.1) (Fig). Rhizopus lipases are synthesized as pre-pro-proteins composed of signal sequence, prosequence and mature lipase sequence (76) (80). The production of Rhizopus lipases has been performed in Escherichia coli [8], in Saccharomyces cerevisiae [9] and in Pichia pastoris [10,11]. Due to either different protein post-translational modifications affecting the glycosylation pattern of these lipases, or to proteolytic cleavage products arising from the mature, the pro- or the pre-pro-enzyme, the lipases derived from recombinant strains showed different molecular mass and different enzymatic properties compared with those from wild strains (75) (81). E. coli as host When E. coli was selected as a host, overexpression of R. oryzae prolipase (ProROL) and mature R. oryzae lipase (mROL) was achieved by using E. coli BL321, the heat-inducible vector pCYTEXP1 and the preOmpA sequence to achieve secretion into the periplasmic space (77) (76). Both fusion proteins induced by a temperature shift to 42  °C were correctly processed and translocated to the periplasmic space, but both mROL and ProROL accumulated as insoluble aggregates. Furthermore, the enzyme activity of mROL damages cells even at low concentrations, whereas the expression of active ProROL has no toxic effects on E. coli and the amount of purified active ProROL (103 U/mL) is larger than that of mROL by a factor of more than 100(77). By in vitro refolding of the insoluble form of mROL, its prosequence was supposed to support the refolding of mROL in vitro (77). R. delemar lipase was also expressed as inactive and insoluble forms in the cytoplasm of E. coli BL21(DE3) using plasmid vector pE T11d (74). In order to achieve soluble production in E. coli, Di Lorenzo et al. (82) used a reductase-deficient host strain, E. coli Origami(DE3), which greatly enhance disulfide bond formation in the E. coli cytoplasm, and pET-11d as expression systems, yielded correctly folded and active mROL and ProROL only in the cytoplasm fraction. Although the specific activities of mROL and ProROL are comparable, the yield of ProROL (166U/mL) was higher than that of mROL (82). S. cerevisiae as host In contrast to E. coli expression system, eukaryotic expression system (eg. P. pastoris, S. cerevisiae) has the advantage of secretion into the cell culture supernatant, avoiding cell disruption, and post-translational modifications, including glycosylation and disulfide bond formation. Takahashi et al. (78) reported the production of the active R. oryzae lipase (ROL) by S. cerevisiae. When the ProROL gene fused with S. cerevisiae ÃŽ ±-factor signal sequence was expressed in S. cerevisiae using UPR-ICL as a promoter, two forms of ROL, ProROL (46 kDa) and r28ROL (35 kDa) having 28 amino acids of the prosequence, were secreted into the culture medium, but the activity of mROL was not detected in both the medium and cells (78). The N-terminal amino acid sequence analysis revealed that the processed form r28ROL was cleaved by Kex2-like protease in S. cerevisiae at the recognition site KR in the prosequence (78). The independent production of two different molecular forms of ROL was succe ssfully obtained using Kex2-engineered strains of S. cerevisiae (83). The amounts of enzyme secreted were estimated as 78.8 mg/l broth for ProROL and 38.3 mg/l broth for r28ROL. These two forms of ROL exhibited distinct properties. ProROL were more thermostable than the processed lipase r28ROL (83), which was also observed in the case of R. niveus lipase expressed in S. cerevisiae (84). The expressed R. niveus lipase activity in the culture supernatant reached 1600 U/mL, a value equivalent to 0.2~0.3 g lipase protein per 1 liter of culture supernatant (72). The role of the prosequence of ROL was further analyzed in S. cerevisiae. When mROL was co-expressed in trans with the prosequence fused to the pre-alpha-factor leader sequence, the activity of mROL was recovered, suggested that the covalent linkage of the prosequence to the mROL was not necessary for the function (85) (86) . From the expression of the ROL mutants with deletions at the N-terminal end of the prosequence together w ith mROL in trans, the residues from 20 to 37 in the prosequence were essential for the secretion, and those from 38 to 57 were essential for the formation of the active ROL and might play a role as an intramolecular chaperone (86). Apart from extracellular expression, R. oryzae lipase was also successfully displayed on the cell surface of S. cerevisiae with a lipase activity of 350.6 U/L (87-89). P. pastoris as host On the contrary to S. cerevisiae expression system when the prolipase gene from R. arrhizus was expressed in P. pastoris, only the mature lipase attached with 28 amino acids of the carboxy-terminal part of the prosequence was secreted in the supernatant (Niu, Li et al. 2006). Prolipase with complete prosequence was nearly found in the supernatant which may be the result that the activity of Kex2-like protease was higher in P. pastoris than in S. cerevisiae (Niu, Li et al. 2006). Interestingly, expression of the prolipase gene from R. chinensis in P. pastors produced two forms of lipase, mature lipase attached with 27 amino acids of the prosequence (r27RCL) and mature lipase (mRCL) (Yu, Wang et al. 2009). At the early stage of fermentation, the recombinant P. pastors produced only r27RCL, which was gradually degraded into mRCL along with the fermentation, probably processed by serine proteases presented in the culture medium (Yu, Wang et al. 2009). For R. oryzae lipase, r28ROL was als o observed to convert to mature lipase gradually incubated at 0-6  °C for a few days (Sayari, Frikha et al. 2005, Hama, Tamalampudi et al. 2006). It has been reported that the prosequence from R. chinensis lipases was more efficient than that from ROL in the role of facilitating the folding and secretion of an active lipase (Yu, Sha et al. 2013). The activity of the chimeric lipase from R. oryzae by replacing the prosequence with that from R. chinensis reached 4050 U/mL, which was 11 fold higher than that of the parent expressed in P. pastoris (Yu, Sha et al. 2013). mROL activity expressed in S. cerevisiae was not detected (Ueda, Takahashi et al. 2002), whereas mROL was functionally expressed and secreted in P. pastoris (Beer, McCarthy et al. 1998) (Minning, Schmidt-Dannert et al. 1998). The expression of mROL was compared under different regulated promoters (Serrano 2002) (Cos, Resina et al. 2005) (Resina, Serrano et al. 2004), i.e. alcohol oxidase 1 promoter (PAOX), glyceraldehyde-3-phosphate dehydrogenase promoter (PGAP) and the formaldehyde dehydrogenase 1 promoter (PFLD1). Expression of the mature form of ROL under the constitutive PGAP promoter resulted in extremely low extracellular lipase levels (Serrano 2002). PAOX is highly induced during growth on methanol, but tightly repressed during growth of the yeast on most common carbon sources such as glucose, glycerol or ethanol, while PFLD1 allows the design of methanol-free culture strategies, being methylamine a less volatile and flammable inducer. Valero and co-authors (Resina, Cos et al. 2005) (Cos, Resina et al. 2005) successfully expressed mROL in P. pastoris under the PFLD1 promoter using methylamine as nitrogen and sorbitol as carbon source. The productivity under PFLD1 promoter are comparable to the classic PAOX system (Resina, Cos et al. 2005) (Cos, Resina et al. 2005). However, limitationsincurrent knowledgein the regulation of methylamine metabolism in P. pastoris limit thedesignof efficientfermentationstrategies using PFLD1 promoter. The presence of multiple integrated copies of a desired expression cassette has been reported to be an important factor in increasing foreign protein production in P. pastoris (Cos, Serrano et al. 2005). The high-level expression of R. chinensis lipase was achieved by optimization of the lipase gene copy number in the host strain P. pastoris (Sha, Yu et al. 2013). Among 1, 3, 5 and 6-copy strains, the maximum lipase activity reached 12,500 U/mL in 5-cope strain, which was 6.2-fold higher than that in 1-copy strain (Sha, Yu et al. 2013). Maximizing gene copy number sometimes resulted in a decreased final productivity yield (Sha, Yu et al. 2013) or a negative effect on cell growth (Cos, Serrano et al. 2005), which indicated stress effects of Rhizopus lipases overexpression upon P. pastoris cells. Recent transcriptomic-based studies (Sha, Yu et al. 2013) (Resina, Bollok et al. 2007) strongly suggest that heterologous overexpression of Rhizopus lipases in P. pastoris indeed triggers the unfolded protein response (UPR). However, co-expression of certain chaperons could relief the cell stress caused by foreign protein overexpression. The productivity of R. chinensis lipase in P. pastoris was improved by co-expression with ERO1p and PDI (Kumar, Jahan et al. 2013) (Sha, Yu et al. 2013). These two chaperons are involved in the protein disulfide bond formation pathway in the endoplasmic reticulum, thus chaperon co-expression contribute to the relief of the protein overexpression stress in recombinant P. pastoris (Kumar, Jahan et al. 2013) (Sha, Yu et al. 2013). Co-expression of unfolded protein response transcriptional factor (Hac1) resulted in about a 3-fold increase in the overall specific productivity of mROL in P. pastoris while the deletion of GAS1, required for the cell wall assembly, achieved 4-fold increase. And the double mutant HAC1/Δgas1 strain yielded about a 7-fold increase (Resina, Maurer et al. 2009). Novel genetic factors enhancing heterologous protei n secretion in P. pastoris have been identified on the basis of transcriptomic analyses (Gasser, Sauer et al. 2007), which give a new insight into engineering P. pastoris for heterologous expression of foreign proteins. Another factor that should be considered for expression optimization is the Mut phenotype. When single copy mROL was expressed in P. pastoris under control of PAOX, the Maximal lipase activity (205U/mL), the specific production rate (qp, 63 U/gbiomass/h ) and the yield of lipase activity per biomass unit (YP/X, 5775U/gbiomass) by Muts strain were 1.37-, 1.29- and 2.34-fold higher than those by Mut+ strain, while the productivity (2246 U/L/h) by Muts strain was 1.34-fold lower than that by Mut+ strain (Cos, Serrano et al. 2005). For R. chinensis prolipase expressed in P. pastoris, Muts strain exibited 1.4-fold higher activity than that of Mut+ phenotype (Yu, Wang et al. 2009). Muts is not as sensitive as Mut+ to high transient methanol concentrations which make the bioprocess easier to control and enables scale up. However, Muts phenotype fermentation results in long induction times with low growth rates (Arnau, Casas et al. 2011). Different strategies have been implemented in the production of R. oryzae lipase under the PAOX promoter in P. pastoris Mut+ phenotype, eg. dissolved oxygen control (DO-stat), methanol limited fed-batch (MLFB, control of the substrate concentration close to zero), methanol non-limited fed-batch (MNLFB, maintenance of the substrate concentration at a constant value), as well as temperature limited fed-batch (TLFB). The MNLFB strategy for ROL expression led to 20–30 h reduction in the production time, a 11-fold higher final lipolytic activity, a 13.6-fold higher productivity and a 10.3-fold higher specific productivity compared to the DO-stat strategy (Minning, Serrano et al. 2001). After optimization of methanol concentration, the activity of Rhizopus chinensis lipase reached 2130 U/mL by maintaining methanol concentration at 1 g/L controlled by an on-line methanol analyzer (Wu, Yu et al. 2011). One of the main drawbacks of P. pastoris Mut+ phenotype is the high oxygen demand i n high cell density cultures using MNLFB strategy (Cos, Serrano et al. 2005). A TLFB strategy was thereafter applied to solve oxygen transfer limitations (Surribas, Stahn et al. 2007). Barrigà ³n et al. (Manuel Barrigon, Luis Montesinos et al. 2013) compared MLFB and MNLFB strategies for the production of mROL expressed in P. pastoris. Yields, productivities and specific production rate in all MLFB conditions were very low. Best results were obtained by MNLFB strategy at methanol set-point of 3 g/L, under which condition maximum lipase activity, productivity and specific productivity reached 280 U/mL, 5406 U/L/h, 102 U/g/h, respectively (Manuel Barrigon, Luis Montesinos et al. 2013) . Due to low specific growth rate of Muts phenotype, strategies to use mixed carbon sources in the induction phase are applied to improve the expression levels of mROL and to increase the productivity of the bioprocess. Glycerol and sorbitol are both frequently used as co-substrates. When glycerol was used as co-substrate, one of the key advantages is its higher ÃŽ ¼ (0.18 h−1 versus 0.02 h−1) compared with sorbitol. However, when the relation ÃŽ ¼Gly per ÃŽ ¼MeOH was larger than 4, a significant decrease of YP/X, volumetric and specific productivity was observed due to the repression of glycerol (Arnau, Casas et al. 2011). The sorbitol as an excellent non-repressive carbon source avoids the severe decrease of the specific production rate and it permits to achieve higher mROL production (Arnau, Ramon et al. 2010) (Ramon, Ferrer et al. 2007). During the induction stage, the ammonium ion released into the fermentation broth has a deep impact on cell growth and protein expression. In our previous study, the impact of NH4+ concentration on the expression of the R. oryzae chimeric lipase in P. pastoris co-expressed with ERO1p and PDI was investigated (Yu, Lu et al. 2013). Analysis of carbon metabolism and energy regeneration pattern revealed that under the optimum NH4+ concentration of 440 mmol/L more carbon source (methanol) was consumed with surged AOX activity and then the higher energy and amino acid precursors demand for recombinant protein synthesis is compensated for by the TCA cycle. Under the optimum NH4+ concentration the lipase activity reached 12,019 U/mL, which was 1.7-fold higher than that without supplement of NH4+ (Yu, Lu et al. 2013). It was also useful for other recombinant protein expression in P. pastoris by addition of inorganic nitrogen source (Kobayashi, Kuwae et al. 2000) (Zhang, Wang et al. 2008) (Yan g and Zhou 2004).