Production of 2,3-butanediol in Saccharomyces cerevisiae by in silico aided metabolic engineering

BACKGROUND: 2,3-Butanediol is a chemical compound of increasing interest due to its wide applications. It can be synthesized via mixed acid fermentation of pathogenic bacteria such as Enterobacter aerogenes and Klebsiella oxytoca. The non-pathogenic Saccharomyces cerevisiae possesses three different 2,3-butanediol biosynthetic pathways, but produces minute amount of 2,3-butanediol. Hence, we attempted to engineer S. cerevisiae strain to enhance 2,3-butanediol production. RESULTS: We first identified gene deletion strategy by performing in silico genome-scale metabolic analysis. Based on the best in silico strategy, in which disruption of alcohol dehydrogenase (ADH) pathway is required, we then constructed gene deletion mutant strains and performed batch cultivation of the strains. Deletion of three ADH genes, ADH1, ADH3 and ADH5, increased 2,3-butanediol production by 55-fold under microaerobic condition. However, overproduction of glycerol was observed in this triple deletion strain. Additional rational design to reduce glycerol production by GPD2 deletion altered the carbon fluxes back to ethanol and significantly reduced 2,3-butanediol production. Deletion of ALD6 reduced acetate production in strains lacking major ADH isozymes, but it did not favor 2,3-butanediol production. Finally, we introduced 2,3-butanediol biosynthetic pathway from Bacillus subtilis and E. aerogenes to the engineered strain and successfully increased titer and yield. Highest 2,3-butanediol titer (2.29 g·l(-1)) and yield (0.113 g·g(-1)) were achieved by Δadh1 Δadh3 Δadh5 strain under anaerobic condition. CONCLUSIONS: With the aid of in silico metabolic engineering, we have successfully designed and constructed S. cerevisiae strains with improved 2,3-butanediol production

Alleviation of carbon catabolite repression in Enterobacter aerogenes for efficient utilization of sugarcane molasses for 2,3-butanediol production

Table S3. Comparison of fed-batch fermentation with EMY-01, EMY-68, EMY-70S, and EMY-70SP using sugarcane molasses

Isolation and characterization of NgRLK1, a receptor-like kinase of Nicotiana glutinosa that interacts with the elicitin of Phytophthora capsici

Elicitins, extracellular proteins from Phytophthora fungi, elicit a hypersensitivity response (HR), including systemic acquired resistance, in some plants. The elicitin capsicein (~10 kDa) was purified by FPLC from culture filtrates of P. capsici. Purified native and recombinant capsicein induced a hypersensitive response in leaves of the non-host plants Nicotiana glutinosa and Brassica rapa subsp. pekinensis. To search for candidate capsicein-interacting proteins from N. glutinosa, a yeast two-hybrid assay was used. We identified a protein interactor that is homologous to a serine/threonine kinase of the plant receptor-like kinase (RLK) group and designated it NgRLK1. The ORF of NgRLK1 encodes a polypeptide of 832 amino acids (93,490 Da). A conserved domain analysis revealed that NgRLK1 has structural features typical of a plant RLK. NgRLK1 was autophosphorylated, with higher activity in the presence of Mn2+ than Mg2+

MicroRNAs Are Indispensable for Reprogramming Mouse Embryonic Fibroblasts into Induced Stem Cell-Like Cells

MicroRNAs play a pivotal role in cellular maintenance, proliferation, and differentiation. They have also been implicated to play a key role in disease pathogenesis, and more recently, cellular reprogramming. Certain microRNA clusters can enhance or even directly induce reprogramming, while repressing key proteins involved in microRNA processing decreases reprogramming efficiency. Although microRNAs clearly play important roles in cellular reprogramming, it remains unknown whether microRNAs are absolutely necessary. We endeavored to answer this fundamental question by attempting to reprogram Dicer-null mouse embryonic fibroblasts (MEFs) that lack almost all functional microRNAs using a defined set of transcription factors. Transduction of reprogramming factors using either lentiviral or piggyBac transposon vector into two, independently derived lines of Dicer-null MEFs failed to produce cells resembling embryonic stem cells (ESCs). However, expression of human Dicer in the Dicer-null MEFs restored their reprogramming potential. Our study demonstrates for the first time that microRNAs are indispensable for dedifferentiation reprogramming

Surgical ciliated cyst of the mandible after orthognathic surgery: a case report with review of the literature

Background : Surgical ciliated cysts, also known as postoperative maxillary cysts or implantation cysts, occur mainly in the posterior maxilla after radical maxillary sinus surgery; they rarely develop in the mandible. They are thought to occur when the sinonasal epithelium is infiltrated by a surgical instrument during surgery or as a result of transplantation of bone or cartilage with respiratory epithelium attached. Case presentation : We report a case in which a surgical ciliated cyst developed in the anterior part of the mandible, presumably as a result of bimaxillary orthognathic surgery and genioplasty performed 24 years earlier. We then review the few similar cases reported in the literature. Conclusion : Surgical ciliated cysts in the mandible are extremely rare, but they could occur after simultaneous surgery on the maxilla and mandible, even decades later. To prevent surgical ciliated cysts in the mandible, we recommend that the surgical instruments, especially the saw blade used during bimaxillary surgery, be new or cleaned and that previously placed plates and screws be removed at an appropriate time

Intradural Extramedullary Tuberculoma Mimicking En Plaque Meningioma

A 24-year-old man with tuberculosis meningitis developed acute paraplegia and sensory disturbances 5 weeks after receiving conventional antituberculous therapy. Magnetic resonance imaging revealed an intradural extramedullary long segmental mass mimicking en plaque meningioma at the T2-T6 vertebrae levels. Prompt surgical decompression was performed. A histology examination of the mass revealed a tuberculoma. After surgery, the patient showed improved motor power and a normal bladder function. Intradural extramedullary tuberculoma of the spinal cord is rare complication of tuberculosis meningitis, which can occur as a response to conventional antituberculous therapy

Removal of Pb(II) from aqueous solution by a zeolite–nanoscale zero-valent iron composite

The effectiveness of nanoscale zero-valent iron (nZVI) to remove heavy metals from water is reduced by its low durability, poor mechanical strength, and tendency to form aggregates. A composite of zeolite and nanoscale zero-valent iron (Z–nZVI) overcomes these problems and shows good potential to remove Pb from water. FTIR spectra support nZVI loading onto the zeolite and reduced Fe0 oxidation in the Z–nZVI composite. Scanning electron micrographs show aggregation was eliminated and transmission electron micrographs show well-dispersed nZVI in chain-like structures within the zeolite matrix. The mean surface area of the composite was 80.37 m2/g, much greater than zeolite (1.03 m2/g) or nZVI (12.25 m2/g) alone, as determined by BET-N2 measurement. More than 96% of the Pb(II) was removed from 100 mL of solution containing 100 mg Pb(II)/L within 140 min of mixing with 0.1 g Z–nZVI. Tests with solution containing 1000 mg Pb(II)/L suggested that the capacity of the Z–nZVI is about 806 mg Pb(II)/g. Energy-dispersive X-ray spectroscopy showed the presence of Fe in the composite; X-ray diffraction confirmed formation and immobilization of Fe0 and subsequent sorption and reduction of some of the Pb(II) to Pb0. The low quantity of Pb(II) recovered in water-soluble and Ca(NO3)2-extractable fractions indicate low bioavailability of the Pb(II) removed by the composite. Results support the potential use of the Z–nZVI composite in permeable reactive barriers

Removal of Pb(II) from aqueous solution by a zeolite–nanoscale zero-valent iron composite

The effectiveness of nanoscale zero-valent iron (nZVI) to remove heavy metals from water is reduced by its low durability, poor mechanical strength, and tendency to form aggregates. A composite of zeolite and nanoscale zero-valent iron (Z–nZVI) overcomes these problems and shows good potential to remove Pb from water. FTIR spectra support nZVI loading onto the zeolite and reduced Fe0 oxidation in the Z–nZVI composite. Scanning electron micrographs show aggregation was eliminated and transmission electron micrographs show well-dispersed nZVI in chain-like structures within the zeolite matrix. The mean surface area of the composite was 80.37 m2/g, much greater than zeolite (1.03 m2/g) or nZVI (12.25 m2/g) alone, as determined by BET-N2 measurement. More than 96% of the Pb(II) was removed from 100 mL of solution containing 100 mg Pb(II)/L within 140 min of mixing with 0.1 g Z–nZVI. Tests with solution containing 1000 mg Pb(II)/L suggested that the capacity of the Z–nZVI is about 806 mg Pb(II)/g. Energy-dispersive X-ray spectroscopy showed the presence of Fe in the composite; X-ray diffraction confirmed formation and immobilization of Fe0 and subsequent sorption and reduction of some of the Pb(II) to Pb0. The low quantity of Pb(II) recovered in water-soluble and Ca(NO3)2-extractable fractions indicate low bioavailability of the Pb(II) removed by the composite. Results support the potential use of the Z–nZVI composite in permeable reactive barriers