The Bifunctional Alcohol and Aldehyde Dehydrogenase Gene, adhE, Is Necessary for Ethanol Production in Clostridium thermocellum and Thermoanaerobacterium saccharolyticum

Thermoanaerobacterium saccharolyticum and Clostridium thermocellum are anaerobic thermophilic bacteria being investigated for their ability to produce biofuels from plant biomass. The bifunctional alcohol and aldehyde dehydrogenase gene, adhE, is present in these bacteria and has been known to be important for ethanol formation in other anaerobic alcohol producers. This study explores the inactivation of the adhE gene in C. thermocellum and T. saccharolyticum. Deletion of adhE reduced ethanol production by \u3e95% in both T. saccharolyticum and C. thermocellum, confirming that adhE is necessary for ethanol formation in both organisms. In both adhE deletion strains, fermentation products shifted from ethanol to lactate production and resulted in lower cell density and longer time to reach maximal cell density. In T. saccharolyticum, the adhE deletion strain lost \u3e85% of alcohol dehydrogenase (ADH) activity. Aldehyde dehydrogenase (ALDH) activity did not appear to be affected, although ALDH activity was low in cell extracts. Adding ubiquinone-0 to the ALDH assay increased activity in the T. saccharolyticum parent strain but did not increase activity in the adhE deletion strain, suggesting that ALDH activity was inhibited. In C. thermocellum, the adhE deletion strain lost \u3e90% of ALDH and ADH activity in cell extracts. The C. thermocellum adhE deletion strain contained a point mutation in the lactate dehydrogenase gene, which appears to deregulate its activation by fructose 1,6-bisphosphate, leading to constitutive activation of lactate dehydrogenase

Increase in Ethanol Yield via Elimination of Lactate Production in an Ethanol-Tolerant Mutant of Clostridium Thermocellum

Large-scale production of lignocellulosic biofuel is a potential solution to sustainably meet global energy needs. One-step consolidated bioprocessing (CBP) is a potentially advantageous approach for the production of biofuels, but requires an organism capable of hydrolyzing biomass to sugars and fermenting the sugars to ethanol at commercially viable titers and yields. Clostridium thermocellum, a thermophilic anaerobe, can ferment cellulosic biomass to ethanol and organic acids, but low yield, low titer, and ethanol sensitivity remain barriers to industrial production. Here, we deleted the hypoxanthine phosphoribosyltransferase gene in ethanol tolerant strain of C. thermocellum adhE*(EA) in order to allow use of previously developed gene deletion tools, then deleted lactate dehydrogenase (ldh) to redirect carbon flux towards ethanol. Upon deletion of ldh, the adhE*(EA) Δldh strain produced 30% more ethanol than wild type on minimal medium. The adhE*(EA) Δldh strain retained tolerance to 5% v/v ethanol, resulting in an ethanol tolerant platform strain of C. thermocellum for future metabolic engineering efforts

NADH-Mediated Gene Expression in Streptococcus pneumoniae and Role of Rex as a Transcriptional Repressor of the Rex-Regulon

Nicotinamide adenine dinucleotides (NAD(H)) play a vital role in various biological processes, including keeping the cellular redox balance. In this study, we investigate the regulatory responses of Streptococcus pneumoniae D39 to NADH and characterize the role of the Rex protein as a transcriptional repressor of the gapN, fba, pncB, adhB2, gap, and adhE genes. Transcriptomic analysis was used to observe the response of S. pneumoniae D39 to NADH. Our microarray studies revealed elevated expression of various genes/operons involved in transport and biosynthesis of niacin (gapN, fba, pncB, adhB2, gap, and adhE). Promoter lacZ-fusion assays and microarray studies with the rex mutant revealed the role of Rex as a transcriptional repressor of gapN, fba, pncB, adhB2, gap, and adhE involved in niacin uptake and biosynthesis, in the presence of NADH. We predict the operator site (5 ' TTGTKAWAAWWTTCACAA-3 ' of Rex in the regulatory regions of Rex-regulated genes that was subsequently validated by promoter mutational experiments

THE IMPLEMENTATION OF PICTURE SEQUENCE TECHNIQUE IN TEACHING VOCABULARY AT THE ELEVENTH GRADE OF THE SECOND SEMESTER OF MA AL-HIKMAH BANDAR LAMPUNG IN THE ACADEMIC YEAR OF 2016/2017

Vocabulary is one of the important language components that need to be taught to support learners in mastering language skill. This research was about teaching and learning vocabulary through picture sequence technique at the eleventh grade of the second semester of MA Al-Hikmah Bandar Lampung in the academic year of 2016/2017. The objectives of this research were to describe teaching learning process in teaching vocabulary through picture sequence technique, to describe the teacher’s problems and students’ problem in teaching and learning process. In this research, the researcher used descriptive qualitative research. The researcher used purposive sampling technique to determine the sample. The researcher chose class XI IPA as sample which consisted of 41 students. In collecting the data, the researcher used three kinds of the instruments, they were: observation, interview and questionnaire. The researcher used three major phases of data analysis, they were: data reduction, data display and conclusion drawing or verification. The results of the research were as follows: first the teaching and learning process by using picture sequence technique was not running well. The classroom atmosphere was not serious in learning process because some of students were noisy. The second, the teacher got some problems, they were: the teacher found difficulties to manage class, used the picture sequence technique takes a lot of time, and the teacher was difficult to increase students’ motivation. The third, the students got some problems in learning vocabulary, they were: the students considered the teacher’s explanation boring, they thought vocabulary learning as knowing the primary meaning of the new words that are gotten only from textbooks or from the teacher. So it was difficult for them to use the words based on the appropriate context, students had difficulty to see these picture clearly and the students were hard to express their ideas and opinions in accordance with the sequence images

Pengaruh Latihan Aktivitas Maze Dan Target Terhadap Keterampilan Teknik Dasar Sepakbola SSB Tunas Jaya Kota Jambi

Based on researcher observation inthe field, showedthat there was stilla lack ofbasicfootballskills in studentsSSBTunasJayaCity ofJambi. This research aimed to observe the effects of maze exerciseactivity and targetactivitytowardbasic technique skill offootball. The hyphotesis of the research were (1)mazeexercise activity gaveinfluence tobasictechnique skillof football. (2) Target exercise activity gave influence to basic technique skill of football. (3) Target exercise activity gave influence to basictechnique skill of football.The population of the researchwasfootballplayerswho joinedonTunasJaya club in Jambi. The active player was about 80 persons.The samplesof the research werejuniorgroupDamount 30persons. It chose because junior group D has 3 month in SSBTunasJayaandtheydidn’t have more skill infootball. The sampling techniquewas done by usingrandom samplingtechniques. The division ofthe sampleinto groupswas done by usingordinally matchingpairing. In techniques of data collection, the researcher usedfootballskill tests toaged10-12 years.  And for the technique of data analysis, the researcher used t testconductedby ttest formula thatrelated to t sample.The results of the research showed that(1)maze exerciseactivitycanincreasethe basic techniques skill of footballinSSBTunasJayaJambi. (2) Target exerciseactivitycan increasethe basictechniques skillof football inSSBTunasJayaJambi. (3) There was no different between maze exercise activity and target activity in increasing basic technique skill of football inSSBTunasJayaJambi. Kata Kunci : Latihan Aktivitas Maze, Latihan Ativitas Target, Keterampilan Sepakbol

PUSAT PENGEMBANGAN KREATIFITAS ANAK JALANAN KOTA MALANG

PUSAT PENGEMBANGAN KREATIFITAS ANAK JALANAN KOTA MALANG Septafian Adhe Permana 0851010028 ABSTRAKSI Anak jalanan merupakan manusia yang masih kecil yang hidup di jalan, lorong, yang berkaitan dengan sepanjang jalan. Mereka sering terlihat berlalu lalang di jalan saat lampu merah. Berpenampilan lusuh, menggunakan kemampuan yang ada untuk bekerja yang menimbulkan kebisingan serta mengganggu ketertiban lalu lintas atau pengguna jalan, tak jarang jauh dari keindahan dan terkadang melakukan pemaksaan kepada penumpang yang tidak memberikan uang receh kepadanya. Faktor yang menyebabkan anak-anak turun ke jalanan untuk bekerja adalah karena tuntutan ekonomi yang dimiliki keluarganya sangat kurang dari kecukupan. Kondisi ini ternyata telah mengusik Pemerintah Daerah untuk membersihkan ketertiban umum. Petugas Keamanan selalu rutin bereaksi untuk membersihkan para anak jalanan dari kota, akan tetapi sepertinya ini bukanlah cara yang efektif karena sama sekali tidak menunjukkan bahwa anak jalanan mulai berkurang. Pusat Pengembangan Kreatifitas Anak Jalanan Kota Malang berguna untuk memberikan keterampilan dan pengembangan kreatifitas terhadap mereka. Sehingga hal ini dapat memajukan tingkat potensi anak jalanan dan mereka juga mendapat peluang kerja sesuai ketrampilan serta bakat yang mereka miliki, khususnya untuk anak jalanan yang berada di Kota Malang. Dalam wadah ini, para anak jalanan mendapatkan pengajaran tentang keterampilan agar mereka dapat memiliki skill yang bermanfaat untuk memenuhi kebutuhan kerja nantinya. Pusat Pengembangan Kreatifitas Anak Jalanan Kota Malang ini merupakan jenis bangunan yang bersifat rumah singgah, bersifat edukasi, serta informative dalam skala kota. Proyek rancangan Pusat Pengembangan Kreatifitas Anak Jalanan Kota Malang mengambil konsep terhadap bangunan yang modern. Yang dimaksud dengan konsep bangunan modern ialah bangunan yang mengandung citra dinamis, estetis, dan inovativ. Kata Kunci : Anak Jalanan, Keterampilan dan Kreatifitas, Moder