Candida Magnoliae Glycerol Yield

Candida Magnoliae Glycerol Yield Abstract Candida magnoliae, isolated from honey comb and Candida glycerinogenes, isolated from natural environment were compared for their potential to produce glycerol from glucose. The highest yield of glycerol was 55% for C. magnoliae and 64.5% for C. glycerinogenes. C. glycerinogenes yields 9% of higher concentration of glycerol than C. magnoliae. For C. magnoliae the optimum conditions were a temperature of 30 °C and a pH of 5. The optimum conditions for C. glycerinogenes were a temperature of 32 °C  and a pH of about 5. The required medium composition for glycerol production was 160 g/L of glucose, 3 g/L of yeast extract, 5 g/L of peptone for C. magnoliae and 230 g/L of glucose, 5 ml/L of corn step liquor, 5-6 g/L of phosphate for C. glycerinogenes. Introduction Glycerol, a simple alcohol which contains three hydroxyl group. Glycerol is also known as glycerine or 1,2,3-Propanetriol. Glycerol has many uses in pharmaceutical, food, paint, cosmetic industries. Glycerol can be used additional fuel in boilers due to its high calorific value. The physiochemical properties and chemical composition of glycerol varies from other fuels. Glycerol is a odourless, colourless, viscous liquid with sweet taste. Glycerol contains three hydrophilic hydroxyl groups which is responsible for its solubility in water. Glycerol has a melting point of 17.9 oC and boiling point of 290 oC. Molecular formula of glycerol is CH2OH-CHOH-CH2OH. Glycerol can be transformed to various value added chemicals such as dihydroxyacetone, succinic acid, citric acid, ethanol, hydrogen etc., Until now the fermentative metabolism of glycerol was being reported in species of bacteria like Citrobacter sp, Enterobacter sp, Lactobacillus sp, Propionibacterium sp, Clostridium and many fungi species. Glycerol has become an abundant carbon source and inexpensive. Glycerol is also produced by yeast fermentation process. Glycerol is obtained as a byproduct during the fermentation of sugar to ethanol using Saccharomyces cerevisiae. Increased glycerol production from monosaccharides can be obtained using yeast fermentation. The production of glycerol in the laboratory is possible by yeast Candida magnoliae and osmotolerant yeast Candida glycerinogenes. Using genetic information, there are new possibilities in the field of fermentation and metabolic engineering. The Overexpression or blocking of genes could potentially can increase yield or productivity. Triose phosphate isomerase is an important enzyme in glycolytic pathway that directs dihydroxyacetone phosphate to glyceraldehyde 3-phosphate. When this triose phosphate isomerase gene was deleted, the mutant is able to achieve higher yield of glycerol. Overexpression of GPD1 gene in yeast increases glycerol production simultaneously increases the accumulation of byproducts such as succinate, acetate, pyruvate etc., This work briefs about the comparison of glycerol production in Candida magnoliae and Candida glycerinogenes. For the design of fermentation process, culture media optimization is an essential step. Many parameters such as phosphate, sulfate, temperature and pH have been found to affect the productivity of glycerol by these microorganisms. Hence these were optimized in prior to other parameters. Materials and Methods Organisms and Media All fermentation procedures were carried out with both C. magnoliae and C. glycerinogenes. C. magnoliae cells were propagated in medium containing 3 g/L of yeast extract, 160 g/L of glucose, 3 g/L of malt extract, 5 g/L of peptone. C. glycerinogenes cells were propagated in medium containing 150 g/L of glucose, 2 g/L of urea and 7 ml/L of corn steep liquor. YEP medium is required to grow C. glycerinogenes and YM medium is required to grow C. magnoliae. The cells were grown until the density reaches 0.2 OD. Fermentation process Fermentations were carried out in 250 ml shake flasks with a working volume of 50 ml. To the working medium 5% (v/v) of C. glycerinogenes and C. magnoliae were inoculated in different flasks. Flask for C. glycerinogenes was incubated at 31 oC and flask for C. magnoliae was incubated at 30 oC for 48 hours. Magnetically stirred 3-l fermentor was used for fermentation with working volume of 1-l. The medium was agitated at 500 rpm and aerated at 2.0 l/min. Analytical methods Glucose concentration was determined using glucose analyzer and glycerol concentration was monitored and confirmed with HPLC using an Aminex HPX 87H column with differential refractive index detector. Other compounds like ethanol, organic acid were analyzed using gas chromatography. Results Candida magnoliae was isolated from honey comb. C. magnoliae can utilize glucose as a carbon source for growth. It strongly ferments glucose to glycerol. This yeast is able to grow in YM medium containing glucose, yeast extract, malt extract and peptone. C. glycerinogenes was isolated from natural environment of high osmotic pressure. Glucose can be used as carbon source by this organism for glycerol production. Optimization Effect of Glucose concentration on glycerol production The effect of different concentration of glucose and glycerol production by C. magnoliae and C. glycerinogenes was determined in 250 ml flask containing 50 ml of medium. For C. magnoliae, concentration of glycerol yield increases when the concentration of glucose was increased from 100 g/l to 200 g/l and for C. glycerinogenes, concentration of glycerol increases when the concentration of glucose ranges between 150 g/L to 250 g/L . Further increase in the concentration of glucose causes a remarkable decrease in the yield. The optimum concentration of glucose for the growth using C. magnoliae was found to be 160 g/L and for C. glycerinogenes, glucose concentration was found to be 230 g/L. Table 1 Effect of different concentration of sulfate on glycerol productivity Glucose (g/L) Glycerol for C. magnoliae (g/L) Glycerol for C. glycerinogenes (g/L) 0 40.9 99.8 100 45.3 104.1 130 49.1 110.3 160 52.6 116.7 190 45.0 120.9 210 41.2 123.2 240 38.5 117.4 270 34.6 106.8 Fig. 1 Effect of initial concentration of glucose in the medium on the production of glycerol by C.magnoliae and C. glycerinogenes based on the amount of glucose consumed. Effect of Phosphate concentration on glycerol production It was found that phosphate is also an important factor in determining the glycerol productivity. A concentration of phosphate between 0 and 2 g/l increases the glycerol yield. Beyond 2 g/l of phosphate the yield of glycerol decreases gradually for C. magnoliae simultaneously the glycerol concentration for C. glycerinogenes decreases beyond 6 g/l of phosphate. Table 2 Effect of different concentration of phosphate on glycerol productivity Phosphate (g/L) Glycerol for C. magnoliae (g/L) Glycerol for C. glycerinogenes (g/L) 0 45.8 76.2 2 46.2 84.1 4 43.8 119.6 6 39.5 123.3 8 37.9 81.3 10 30.3 47.2 Fig. 2 Effect of initial concentration of phosphate on the production of glycerol in the medium by C.magnoliae and C. glycerinogenes based on the amount of consumed glucose. Effect of Temperature on glycerol production Using shake-flask culture, the optimum temperature was determined for glycerol production. The concentration of glycerol varies with temperature from 26 oC to 34 oC. The yield of glycerol increases till 30 oC for C.magnoliae and beyond this temperature the yield decreases. Similarly for C.glycerinogenes concentration of glycerol decreases beyond 32 oC. Table 3 Effect of temperature on glycerol productivity Temperature (oC) Glycerol for C. magnoliae (g/L) Glycerol for C. glycerinogenes (g/L) 26 68.6 79.6 28 73.2 101.1 30 77.3 114.2 32 74.1 130.4 34 70.5 125.7 Fig. 3 The temperature significantly affected the production of glycerol by à ¢-  C.magnoliae and à ¢-   C.glycerinogenes based on the amount of glucose consumed. Effect of pH on glycerol production Batch experiments were done to determine the effect of pH on glycerol production from C.magnoliae and C.glycerinogenes. At acidic pH the yield was found to be low. Between the pH 4 and 6, the production of glycerol was not significantly affected. At pH 5 there was a significant increase in the growth rate and glycerol production. Finally the optimum pH was found to be 5 for both the organisms. Table 4 Effect of pH on glycerol productivity pH Glycerol for C. magnoliae (g/L) Glycerol for C. glycerinogenes (g/L) 3.0 113.4 3.5 4.0 60.3 120.2 4.5 75.9 5.0 80.1 134.7 5.5 76.0 6.0 66.2 127.1 6.5 7.0 103.5 Fig. 4 Effect of initial pH of medium on production of glycerol by à ¢-  C. magnoliae and à ¢-   C. glycerinogenes based on the amount of glucose consumed. Discussion Glycerol yield by microbial fermentation of glucose using S. cerevisiae is less than 50%(w/w). But newly discovered C. magnoliae and osmotolerent yeast C. glycerinogenes produced glycerol in higher concentration compared with S. cerevisiae. 64.5% of glycerol obtained from C. glycerinogenes and C. magnoliae yields 55% of glycerol after recovery. Therefore C. glycerinogenes gives 9% more yield when compared to C. magnoliae. Increase in glycerol by C. glycerinogenes is due to overexpression of GPD1 gene. Also glycerol concentration increases in C. magnoliae when GPD1 gene overexpressed. But it also accumulated higher amount of byproducts such as acetate, succinate, pyruvate and acetoin. Many parameters such as temperature, pH, phosphate and glucose were found to affect the glycerol productivity by C. magnoliae and C. glycerinogenes. The optimum concentration of glucose for C. magnoliae was found to be 160 g/L and for C. glycerinogenes it was found to be 230 g/L, beyond these concentration range the yield of glycerol decreases. 2 g/L of phosphate was estimated as optimum concentration for glycerol production by C. magnoliae and similarly for C. glycerinogenes, maximum glycerol was obtained at 6 g/L of phosphate. 28 oC – 32 oC of temperature favors the good growth of cells and better glycerol production. Thus the optimum temperatures was declared to be 30 oC for C. magnoliae and 32 oC for C. glycerinogenes. Finding different species other than S. cerevisiae will give new knowledge, beyond physiological effect of glycerol production.

Wise Blood Character Annalysis- Enoch Emery

Wise Blood Character Evaluation Essay: Enoch Emery In the book â€Å"Wise Blood† by Flannery O’Connor, Enoch Emery is first introduced into the story in the third chapter when the main character. Emoch runs into the main character Hazel motes on the street in a crowd of people surrounding a man selling potato peelers. Enoch is an eighteen years old and is described as a â€Å"damp haired pimpled boy† with â€Å"yellow hair and a fox-shaped face. †(pg. 5) We are quickly shown his lack of intelligence by his terrible grasp on the English language and him only laughing after being mocked by the peeler sales man.Enoch is an impulsive rude boy, who just does and says what he wants not thinking of the consequences of his actions. He makes new enemies everywhere. Mocking and snickering at waitresses, making inappropriate comments, getting himself called names like â€Å"son of a bitch. † More than anything he is just simple minded. He does these things bec ause he does not think of consequences. He says there are no friendly people in the city, but he probably is just not giving anyone a good reason to be friendly towards him. Enoch is instantly drawn towards Hazel Motes by something he calls â€Å"wise blood. He claims he inherited wise blood from his father and that it tells him what to do and where to go. Enoch believes that his wise blood will come in handy to Hazel in his mission to start â€Å"The Church Without Christ. † Hazel just sees Enoch as an insane obsessive teenage boy though and wants nothing to do with him. Enoch tries to tag along with Hazel but is rejected by him immediately. However, soon after Hazel got rid of Enoch, he found himself having to go to his work at the city zoo to fetch him. Thinking that Enoch knew where to find the blind preacher and his daughter, Hazel decided to follow him.Before Enoch brought Hazel to the preacher he had to show Hazel what he thought was a huge secret. One he could only t ell someone who he felt was the chosen one to share it with. This person had to be someone who he felt was pure and not from the city. He immediately knew it to be Hazel and felt he had to find a way to share what he knew with him. Enoch took Hazel to a museum that was through the woods from his work, telling Hazel that he would not tell him where the preacher lived unless Hazel saw what Enoch wanted to show him.When they got inside Enoch told Hazel to be quiet as to not wake the guard who did not like Enoch very much. When they finally arrived where Enoch wanted to bring Hazel it turned out that what he was showing Hazel was a three foot tall shrunken man in a glass case. â€Å"See theter notice,† Enoch said in a church whisper, pointing to a type written card at the man’s foot, â€Å"it says he was once as tall as you or me. Some A-rabs did it to him in six months. † He turned his head cautiously to see Hazel Motes. † (pg. 94) Enoch believed that this wa s the new Christ that Hazel was searching to find for his Church Without Christ.When Hazel saw this however he wanted nothing to do with the shrunken man and stormed out. Although Enoch had told Hazel that he knew where the two of them lived it was a lie. However he went ahead anyway with Hazel, trying to rely on his wise blood to guide him. In the end they ended up not finding the blind preaches house, only filling Hazel with more hatred towards Enoch. Enoch’s blood was telling him that Hazel needed the shrunken man for his church without Christ so he knew he had to steal it from the museum.Enoch went to the museum with shoe polish covering his face and hands so that if anyone were to see him they would suspect a colored man and not him. Enoch managed to steal the shrunken man without any witnesses. Enoch brought the man home and put him carefully away, afraid to even look at it. He suspected that he would soon be rewarded for his actions. So he sat and waited, but nothing h appened. Enoch waited longer, but still nothing happened. Finally he gave up, slamming the door to the cabinet the shrunken man was in, smashing it’s face.Since nobody saw him steal the shrunken man he could still possibly be a suspect so Enoch put on a hat and a fake beard and headed off to the address that Sabbath Hawks gave him to find Hazel Motes. When he got to the house Sabbath answered Hazel’s door saying that Hazel was ill and persuaded him to leave the package with her, not knowing that it was the shrunken man inside. â€Å"My man is sick today and sleeping,† she said, â€Å"because he didn’t sleep none last night. What you want? † â€Å"This is for him, it ain’t for you,† Enoch said, handing her the wet bundle. A friend of his gave it to me to give to him. I don’t know what’s in it. † â€Å"I’ll take care of it,† she said. â€Å"You needn’t to worry none† (pg. 183) Enoch left s lamming the door and despite Enoch’s request not to open it and to deliver it directly to Hazel she brought it to the bathroom and started to tear open the package, despite Enoch’s wishes of her. Later Enoch discovers a line of children waiting to shake hands with a gorilla named Gonga, which is really just a man in a gorilla suit. Enoch, being the simple minded fool that he is, gets in line with all the small children.The children who are there to see a movie staring Gonga not just to shake hands with a fake gorilla, but Enoch wanted nothing to do with the movie, only wanting to shake the man’s hand and trying each time to strike conversation but then being pushed aside. The man in the Gonga suit ends up telling Enoch to go to hell, this being the first time that anyone from the city reached out their hand to Enoch and then this being the reaction to potential conversation, Enoch was immediately crushed and storms off.Later Enoch is in a diner and reads that Go nga is going to be at another local theater so he tracks him down. Enoch climbs into the back of the van that Gonga arrived in, hid, and waited for them to leave. Once they finished they loaded up the van and left. Enoch stayed in the van until they came to a stop at a crossing where he jumped out holding the Gonga suit and ran off to the woods where he buries his human clothes and sets off into the night ready to meet people as his new identity of Gonga.Overjoyed Enoch feels that he is finally being rewarded as he should be. Enoch walks down to the edge of the road where he finds a man and woman sitting on a rock over looking the city. As he tries to greet them they scream and run off. Enoch’s hands drop to his sides and he is left alone in the dark looking out over the city. This is the last we ever hear from him for the rest of the story. One of Enoch’s main roles is â€Å"Wise Blood† is comic relief. He is pretty much the only character to bring humor to the story throughout the book.He is constantly making bad decisions and always trying to insult people everywhere he goes. He is also where the title of the story comes from. Enoch is the only character in the book that actually claims to have â€Å"wise blood,† which he claims he inherited from his father who also had it. Enoch is clueless in life and the only important decisions he ever makes are ones he feels like he has no choice but to make since he feels his blood is telling him what to do, and it is almost impossible for him to ignore what the wise blood wants.

The Scope and Limitations of the Fraud Examination

On October 5th, 2010, the fraud examination unit received an assignment from the owner of school regarding to the possibility of fraud disbursements occurred in his school. He suspected that someone is illegally and intentionally stealing money from his school. Based on his suspicion, a fraud examination was conducted by collecting information, reviewing & analyzing relevant records, and applying detecting methods. The objective of my examination was to determine the possibility of misappropriation of school’s fund, identify the fraudulent schemes, and help the school to figure out who was behind the fraud. I reviewed school’s bank accounts, checkbooks, and personnel files. After obtaining sufficient documentation, I found that the school funds were intercepted and converted to Mr Bob Daniel, who is the supervisor in school. He was authorized to control financial and the process of check’s preparation and issuance. Since none of financial supervisor reviewed his works, he misappropriated checks and fraudulently affixes the signature of an authorized maker. His forged maker scheme was executed through following steps: – Start scheme with 2 shell companies Prepare fraud checks and forge the signature of school’s owner – Deliver checks to the bank – Fraud checks were payable to his shell companies and his account. He concealed his theft by using high activity accounts. Since tracking records on the high active accounts would not expose the fraud easily, he successfully embezzled approximately $400,000 from school’s funds. I applied detection methods to support my investigation as follows: – Tracking on the vendor expenditures: it would increase compared to prior expenses. Detect on the process of check’s preparation and issuance: Bob Daniel was an important part in the entire process. Also, none of supervisor checked his works. – Examine the personnel records: a review of Bob Daniel showed that rationalization would be a major motivation for his theft – Obtaining confession: he confessed his theft as the result of feeling guilty. In conclusion, this report reflects that Bob Daniel, the school financial supervisor, committed a check tampering to misappropriate approximately $400,000. A full review of internal controls should be conducted to detect and prevent more fraudulent schemes.

Informative Speech on Dreams Essay - 1666 Words

To inform: To inform my audience on dreams Intro: Can you remember the last dream you had? Maybe you could fly or were falling down an endless dark tunnel. Perhaps you were awakened by a horrific dream in the middle of the night. If any of you are like me you experience vivid dreams. I would like to start by share one of my dream experiences with you all. About two months ago I had a dream that my boyfriend was cheating on me, now I know all you girls and guys can relate to this. I wake up in the morning with my dream still fresh on my mind and in a horrible mood. I am furious at my boyfriend, and for what reason? He hasn’t even done anything, but the thing is in my mind he has, because my dream is so vivid and real to me. So guys maybe†¦show more content†¦Dreams have been around as long as the first civilization came to be and have been a normal part of human existence. One third of your life is spent sleeping, and of that third, on average you will have spent a total of about six years of it dreaming. Mos t people dream on average two hours every night, but you can have anywhere from four to seven dreams in one night. According to research, the most common setting for a dream is in your own house. In our dreams we can do anything we want and be whoever we want to be. Our dreams are an escape from reality. While we dream we are unable to control our actions and choose our surroundings. We let our minds take over. Sometimes dreams can be understood in the context of repressed thoughts. Dreaming serves as an outlet for those thoughts and impulses we repress during the day. When we go to sleep at night and slip into our dream state, we feel liberated and behave in a manner that we do not allow ourselves to in our everyday life. Visions and ide s can come from your dreams. 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