Expression, thermal stability modification and application in PHB degradation of polyhydroxyalkanoate depolymerase from Thermomonospora umbrina / 生物工程学报

Polyhydroxyalkanoate depolymerase (PHAD) can be used for the degradation and recovery of polyhydroxyalkanoate (PHA). In order to develop a PHAD with good stability under high temperature, PHAD from Thermomonospora umbrina (TumPHAD) was heterelogously expressed in Escherichia coli BL21(DE3). At the same time, a mutant A190C/V240C with enhanced stability was obtained via rational design of disulfide bonds. Characterization of enzymatic properties showed that the mutant A190C/V240C had an optimum temperature of 60 ℃, which was 20 ℃ higher than that of the wild type. The half-life at 50 ℃ was 7 hours, at 50 ℃ which was 21 times longer than that of the wild type. The mutant A190C/V240C was used for the degradation of polyhydroxybutyrate (PHB), one of the typical PHA. At 50 ℃, the degradation rate of PHB being treated for 2 hours and 12 hours was 2.1 times and 3.8 times higher than that of the wild type, respectively. The TumPHAD mutant A190C/V240C obtained in this study shows tolerance to high temperature resistance, good thermal stability and strong PHB degradation ability, which may facilitate the degradation and recovery of PHB.

Vibrio sp. ArtGut-C1, a polyhydroxybutyrate producer isolated from the gut of the aquaculture live diet Artemia (Crustacea)

BACKGROUND: Vibrio species display variable and plastic fitness strategies to survive and interact with multiple hosts, including marine aquaculture species that are severely affected by pathogenic Vibrios. The culturable Vibrio sp. strain ArtGut-C1, the focus of this study, provides new evidence of such phenotypic plasticity as it accumulates polyhydroxybutyrate (PHB), a biodegradable polymer with anti-pathogen activity, particularly in the marine larviculture phase. The strain was isolated from the gut of laboratory-reared Artemia individuals, the live diet and PHB carrier used in larviculture. Its main phenotypic properties, taxonomic status and genomic properties are reported based on the whole-genome sequencing. RESULTS: Vibrio sp. ArtGut-C1 yielded 72.6% PHB of cells' dry weight at 25 C. The genomic average nucleotide identity (ANI) shows it is closely related to V. diabolicus (ANI: 88.6%). Its genome contains 5,236,997- bp with 44.8% GC content, 3,710 protein-coding sequences, 96 RNA, 9 PHB genes functionally related to PHB metabolic pathways, and several genes linked to competing and colonizing abilities. CONCLUSIONS: This culturable PHB-accumulating Vibrio strain shows high genomic and phenotypic variability. It may be used as a natural pathogen biocontrol in the marine hatchery and as a potential cell factory for PHB production.

Improvement of polyhydroxybutyrate production by deletion of csrA in Escherichia coli

BACKGROUND: Poly-3-hydroxybutyrate (PHB) can be efficiently produced in recombinant Escherichia coli by the overexpression of an operon (NphaCAB) encoding PHB synthetase. Strain improvement is considered to be one of critical factors to lower the production cost of PHB in recombinant system. In this study, one of key regulators that affect the cell growth and PHB content was confirmed and analyzed. RESULT: S17-3, a mutant E. coli strain derived from S17-1, was found to be able to achieve high cell density when expressing NphaCAB with the plasmid pBhya-CAB. Whole genome sequencing of S17-3 revealed genetic alternations on the upstream regions of csrA, encoding a global regulator cross-talking between stress response, catabolite repression and other metabolic activities. Deletion of csrA or expression of mutant csrA resulted in improved cell density and PHB content. CONCLUSION: The impact of gene deletion of csrA was determined, dysfunction of the regulators improved the cell density of recombinant E. coli and PHB production, however, the detail mechanism needs to be further clarified.

Evaluation of short-chain-length polyhydroxyalkanoate accumulation in Bacillus aryabhattai

Abstract This study was focused on the polyhydroxybutyrate (PHB) accumulation property of Bacillus aryabhattai isolated from environment. Twenty-four polyhydroxyalkanoate (PHA) producers were screened out from sixty-two environmental bacterial isolates based on Sudan Black B colony staining. Based on their PHA accumulation property, six promising isolates were further screened out. The most productive isolate PHB10 was identified as B. aryabhattai PHB10. The polymer production maxima were 3.264 g/L, 2.181 g/L, 1.47 g/L, 1.742 g/L and 1.786 g/L in glucose, fructose, maltose, starch and glycerol respectively. The bacterial culture reached its stationary and declining phases at 18 h and 21 h respectively and indicated growth-associated PHB production. Nuclear Magnetic Resonance (NMR) spectra confirmed the material as PHB. The material has thermal stability between 30 and 140 °C, melting point at 170 °C and maximum thermal degradation at 287 °C. The molecular weight and poly dispersion index of the polymer were found as 199.7 kDa and 2.67 respectively. The bacterium B. aryabhattai accumulating PHB up to 75% of cell dry mass utilizing various carbon sources is a potential candidate for large scale production of bacterial polyhydroxybutyrate.

Poly-hydroxybutyrate (PHB) production by Halomonas elongata BK AG 18 indigenous from salty mud crater at central Java Indonesia

Aims: Bioplastic is a biodegradable polymer produced by particular microorganism as a secondary metabolite. Some halophilic bacteria belonging to Halomonas genus have been reported to be a potential of polyhydroxybutyrate (PHB) producer. This study aims to explore the potential of an indigenous halophilic bacterial isolate, H. elongata BK-AG18, as bioplastic producer. The indication as bioplastics producer was evaluated by growing in nile red-containing medium and bacterial colonies displayed bright orange fluorescent under ultraviolet light. Methodology and results: Bioplastic production by H. elongata BK-AG 18 was achieved using modified glucosecontained High Medium (HM) after incubated in a rotary shaker for 22 h, 37 °C, 150 rpm. The bioplastic was extracted with chloroform and sodium hypochlorite (1:1) and precipitated in methanol. The highest yield of bioplastic production was 21.36% of the dried bacterial cell weight. The structural characterizations of the bioplastics using Fourier transformed infrared (FTIR), 1H and 13C nuclear magnetic resonances (NMR) spectroscopies showed high similarity to the spectral pattern of poly-hydroxybutyrate (PHB). Further characterization using differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) revealed that the decomposition and melting temperature at 266 °C and 166.5 °C of the PHB, respectively. The result of PHB has a low degree of crystallinity (9.5%) that close to the rubber-like polymer. Conclusion, significance and impact of study: This study revealed the high potential of H. elongata BK-AG 18 as PHB producer with high mechanical properties.

A purple non-sulfur bacterium producing polyhydroxybutyrate and the conserved region of pha synthase gene / Bactéria púrpura não sulfurosa produtora de polihidroxibutirato e a região conservada do gene pha sintase

This study aimed to screen purple non-sulfur bacteria capable of accumulating granules or polyhydroxybutyrate (PHB) inside the cells, identify the potent strain, assay the enzyme or PHA synthase, and compare the PHB synthase gene with that of related strains. A total of 58 strains of purple non-sulfur bacteria were isolated from 108 samples of chicken feces in the chicken-egg farm of the Department of Animal Science, Faculty of Natural Resources at Prince of Songkla University, Hat Yai, Thailand. After cultivating the bacteria in glutamate malate (GM) medium without added glutamic acid under light (3,000 Lux) at 35oC for 5 days, the intracellular biopolymer granules of the bacteria were observed by using a Confocal Laser Scanning Microscope (CLSM) with excitation and emission wavelength of 530 and 605 nm, respectively. Gas chromatography (GC) was carried out for quantitative analysis of PHB. There were five strains, CH12, CH52, CH72, CH90 and CH92, showed biopolymer granules under CLSM, and accumulated PHB 5, 1.7, 1.5, 1.4 and 1.8% (w w-1) of the cell dry weight (CDW), respectively. The 16S rDNA sequence analysis of CH12 strain showed a high homology of 100% correlation to that of Rhodopseudomonas palustris strain NCIB8288. Regarding the taxonomic characteristics and 16S rDNA sequence analysis, CH12 strain was identified as Rps. palustris NCIB8288. The PHA synthase activity of the crude extract from CH12 strain was 25 units/mL. The conserved regions could be aligned and selected among 5 strains of Rhodopseudomonas palustris (strains BisA53, TIE-1, CGA009, HaA2 and BisB18). The purified PCR product was obtained for further studies.

Este estudo teve como objetivo rastrear bactérias púrpuras não sulfurosas capazes de acumular grânulos ou polihidroxibutirato (PHB) dentro das células, identificar a estirpe potente, ensaiar a enzima ou PHA sintaxe, e comparar com o gene PHB sintase com aquele de estirpes relacionadas. Um total de 58 estirpes de bactérias púrpuras não sulfurosas foram isoladas a partir de 108 amostras de fezes de galinhas na granja produtora de ovos do Departamento de Ciência Animal, Faculdade de Recursos Naturais da Universidade Prince of Songkla, Hat Yai, Tailândia. Depois de cultivar as bactérias em um substrato de glutamato/malato (GM), sem ácido glutâmico adicionado, sob luz (3000 lux) a 35 ºC durante 5 dias, os grânulos de biopolímeros intracelulares das bactérias foram observados utilizando um microscópio confocal (do inglês Confocal Laser Scanning Microscope - CLSM) com comprimentos de onda de excitação e emissão de 530 e 605 nm, respectivamente. A cromatografia gasosa (do inglês Gas chromatography - GC) foi realizada para uma análise quantitativa de PHB. Havia 5 estirpes, CH12, CH52, CH72, CH90 e CH92, que mostraram grânulos biopoliméricos quando submetidos ao CLSM, e PHB-5 acumulado de 1.7, 1.5, 1.4 and 1.8% (w w-1) do peso celular seco (do inglês cell dry weight - CDW), respectivamente. A análise da sequência do rDNA 16S da estirpe CH12 demonstrou uma alta correlação de homologia de 100% para aquela da estirpe NCIB8288 da Rhodopseudomonas palustris. Em relação às características taxonômicas e da análise da sequência do rDNA 16S, a estirpe CH12 foi identificada como Rps. palustris NCIB8288. A atividade da PHA sintase do extrato bruto da estirpe CH12 foi de 25 unidades/mL. As regiões conservadas puderam ser alinhadas e selecionadas entre 5 estirpes de Rhodopseudomonas palustris (BisA53, TIE-1, CGA009, HaA2 e BisB18). O produto purificado da reação em cadeia da polimerase - PCR foi obtido para estudos futuros.

Microbial polyhydroxybutyrate production by using cheap raw materials as substrates.

Bioplastic, Polyhydroxybutyrate (PHB) is well known for it’s environmental friendliness and complete decomposition into water and carbon dioxide by microorganisms. The main drawback of PHB commercialization is it’s high production cost which is 10 times higher than that of synthetic plastic. So, the present research work mainly focussed on the fermentative production of PHB by Bacillus amyloliquifaciens and Nocardiopsis potens using low cost raw materials like Molasses, wheat bran, rice bran, ragi bran, jambul seed powder, orange peel and whey as substrates. Bacillus amyloliquifaciens and Nocardiopsis potens gives maximum PHB 16.5 µg/ ml and 26.8 µg/ ml respectively in the medium containing molasses and wheat bran as a substrates. Further the functional groups of extracted PHB were confirmed by Fourier transform infrared spectroscopy.

Extraction and Characterization of Polyhydroxybutyrates (PHB) from Bacillus thuringiensisKSADL127 Isolated from Mangrove Environments of Saudi Arabia

ABSTRACTPolyhydroxybutyrate (PHB) is a renowned biodegradable plastic that do not release any toxins or residues in the environment like petroleum based plastics. In the present study, 50 bacteria isolated from mangrove niche, Saudi Arabia, were screened for maximum PHB production. All the 50 strains showed positive for PHB production, of which one strain showed maximum of 137 mgL-1. The most PHB accumulated bacterium was selected and identified asBacillus thuringiensis KSADL127, based on phenotypic characterization and 16S rRNA sequence analysis. Characterization of extracted PHB was carried out by FT-IR, NMR, UV spectroscopy, DSC, TGA, and LC-MS, which later confirmed the presence of intracellular accumulated polymer and substantiated as PHB.

Agrowaste-based Polyhydroxyalkanoate (PHA) production using hydrolytic potential of Bacillus thuringiensis IAM 12077

The study identified the innate enzymatic potential (amylase) of the PHB producing strain B.thuringiensis IAM 12077 and explored the same for cost-effective production of PHB using agrowastes, eliminating the need for pretreatment (acid hydrolysis and/or commercial enzyme). Comparative polyhydroxyalkanoate (PHA) production by B. thuringiensis IAM 12077 in biphasic growth conditions using glucose and starch showed appreciable levels of growth (5.7 and 6.8 g/L) and PHA production (58.5 and 41.5%) with a PHA yield of 3.3 and 2.8 g/L, respectively. Nitrogen deficiency supported maximum PHA yield (2.46 g/L) and accumulation (53.3%). Maximum growth (3.6 g/L), PHB yield (2.6 g/L) and PHA accumulation (72.8%) was obtained with C:N ratio of 8:1 using starch as the carbon source (10 g/L). Nine substrates (agro and food wastes) viz. rice husk, wheat bran, ragi husk, jowar husk, jackfruit seed powder, mango peel, potato peel, bagasse and straw were subjected to two treatments- acid hydrolysis and hydrolysis by innate enzymes, and the reducing sugars released thereby were utilized for polymer production. All the substrates tested supported comparable PHB production with acid hydrolysis (0.96 g/L-8.03 g/L) and enzyme hydrolysis (0.96 g/L -5.16 g/L). Mango peel yielded the highest PHB (4.03 g/L; 51.3%), followed by jackfruit seed powder (3.93 g/L; 29.32%). Varied levels of amylase activity (0.25U-10U) in all the substrates suggested the enzymatic hydrolysis of agrowastes.

New PHA products using unrelated carbon sources

Polyhydroxyalkanoates (PHA) are natural polyesters stored by a wide range of bacteria as carbon source reserve. Due to its chemical characteristics and biodegradability PHA can be used in chemical, medical and pharmaceutical industry for many human purposes. Over the past years, few Burkholderia species have become known for production of PHA. Aside from that, these bacteria seem to be interesting for discovering new PHA compositions which is important to different industrial applications. In this paper, we introduce two new strains which belong either to Burkholderia cepacia complex (Bcc) or genomovar-type, Burkholderia cepacia SA3J and Burkholderia contaminans I29B, both PHA producers from unrelated carbon sources. The classification was based on 16S rDNA and recA partial sequence genes and cell wall fatty acids composition. These two strains were capable to produce different types of PHA monomers or precursors. Unrelated carbon sources were used for growth and PHA accumulation. The amount of carbon source evaluated, or mixtures of them, was increased with every new experiment until it reaches eighteen carbon sources. As first bioprospection experiments staining methods were used with colony fluorescent dye Nile Red and the cell fluorescent dye Nile Blue A. Gas chromatography analysis coupled to mass spectrometry was used to evaluate the PHA composition on each strain cultivated on different carbon sources. The synthesized polymers were composed by short chain length-PHA (scl-PHA), especially polyhydroxybutyrate, and medium chain length-PHA (mcl-PHA) depending on the carbon source used.

New PHA products using unrelated carbon sources

Polyhydroxyalkanoates (PHA) are natural polyesters stored by a wide range of bacteria as carbon source reserve. Due to its chemical characteristics and biodegradability PHA can be used in chemical, medical and pharmaceutical industry for many human purposes. Over the past years, few Burkholderia species have become known for production of PHA. Aside from that, these bacteria seem to be interesting for discovering new PHA compositions which is important to different industrial applications. In this paper, we introduce two new strains which belong either to Burkholderia cepacia complex (Bcc) or genomovar-type, Burkholderia cepacia SA3J and Burkholderia contaminans I29B, both PHA producers from unrelated carbon sources. The classification was based on 16S rDNA and recA partial sequence genes and cell wall fatty acids composition. These two strains were capable to produce different types of PHA monomers or precursors. Unrelated carbon sources were used for growth and PHA accumulation. The amount of carbon source evaluated, or mixtures of them, was increased with every new experiment until it reaches eighteen carbon sources. As first bioprospection experiments staining methods were used with colony fluorescent dye Nile Red and the cell fluorescent dye Nile Blue A. Gas chromatography analysis coupled to mass spectrometry was used to evaluate the PHA composition on each strain cultivated on different carbon sources. The synthesized polymers were composed by short chain length-PHA (scl-PHA), especially polyhydroxybutyrate, and medium chain length-PHA (mcl-PHA) depending on the carbon source used.

Production and characterization of poly-3-hydroxybutyrate from crude glycerol by Bacillus sphaericus NII 0838 and improving its thermal properties by blending with other polymers

The aim of this work was to study the production of poly-3-hydroxybutyrate (PHB) under nitrogen limited conditions by Bacillus sphaericus NII 0838 using crude glycerol from biodiesel industry as sole carbon source. Effect of various process parameters on PHB production such as glycerol concentration, inoculum size and pH of the medium were optimized. Characterization of extracted PHB was carried out by FT-IR, ¹H and 13C NMR. Results showed that the bacterial culture accumulated about 31 percent PHB in crude glycerol medium. The extracted PHB was blended with other polymers to improve its physical characteristics. The thermal properties of the polymer like melting temperature (Tm) and heat of fusion (ΔHf) were determined using DSC.

Avaliação mecânica da placa de compósito de poli-hidroxibutirato e hidroxiapatita em modelos ósseos de gato / Evaluation of the mechanics of of polyhydroxybutyrate and hydroxyapatite composite plates in bone models of cat

Avaliou-se mecanicamente o compósito de poli-hidroxibutirato 70 por cento e hidroxiapatita 30 por cento na forma de placas para fixação óssea. Foram utilizadas 15 placas do compósito com 60mm de comprimento por 10mm de largura e espessura variando de 3mm no centro a 5mm nas extremidades, com seis orifícios. As placas foram fixadas em um modelo acrílico de fêmur de gato, e o conjunto foi submetido aos testes de flexão com quatro pontos, compressão axial e torção, empregando-se como referência microplacas de aço ASTM-F138 2,0mm. As médias das forças máximas nos testes de flexão e de compressão foram, respectivamente, de 323,20N e 617,70N, para as placas de compósito, e de 352,33N e 547,70N, para as placa de aço. No teste de torção, as médias dos torques máximos foram de 1,01Nm para as placas de compósito e de 1,15Nm para as placas de aço. Não houve diferença estatística entre as placas de compósito e de aço. O comportamento físico do material foi diferente, pois as placas de compósito se romperam e as de aço apenas se deformaram, revelando baixa ductilidade das placas de compósito.

Fixation bone plates made of 70 percent polyhydroxybutyrate and 30 percent hydroxyapatite composite were mechanically evaluated. The fifteen composite plates employed presented six holes and measured 60 x 10mm, length and width, respectively, with thickness ranging from 3 to 5mm according to the region. The plates were fixed in acrylic models of cat femur, then were subjected to tests of four-points bending, axial compression, and torsion, using as reference 2mm stainless steel plates. The means of the maximum force in flexion and compression tests were, respectively, 323.20N and 617,70N for the composite plates and 352.33N and 547.70N for the steel plates. In the torsional test, the means of torque were 1.01Nm for the composite plates and 1.15Nm for steel plates. There were no statistical differences between the plates of composite and steel. The physical behavior of the material was different once the composite plates broke up while the stainless steel ones only presented deformation, revealing the low ductility of the composite plates.

Production of PHB (polyhydroxybutyrate) by rhodopseudomonas palustris KU003 and rhodobacter capsulatus KU002 under phosphate limitation.

Nine bacterial species of purple non sulphur bacteria were isolated from Warangal district of South India from leather industry effluents which included Rhodopseudomonas palustris, R.rutila, R.acdiophila, Rhodopila globiformis, Rhodospirillum rubrum, Rsp.photometricum, Rhodobacter sphaeroides, Rb.capsulatus, Rhodobacter sp and Rhodocyclus gelatinosus were isolated. Among these Rhodopseudomonas palustris KU003 and Rhodobacter capsulatus KU002 were selected for the production of Polyhydroxybutyrate (PHB). The extracted PHB was characterized by IR and NMR spectral analysis. Effect of nutrient limitation in the form of phosphate KH2PO4 was tested to enhance the production of the polymer. Maximum yield of PHB was recorded at a concentration of 280 mg/L of KH2PO4 in Rps.palustris while no significant increase in the production of the polymer was observed in Rb.capsulatus.

Emprego experimental da placa de compósito poli-hidroxibutirado/hidroxiapatita na fixação femoral em gatos / Experimental employment of polyhydroxybutyrate/hydroxyapatite composite plates in femoral fixation in cats

Avaliou-se o compósito de poli-hidroxibutirado (PHB) 70 por cento e hidroxiapatita (HA) 30 por cento na forma de placas para fixação óssea em gatos. Foram usadas placas do compósito com 60mm de comprimento por 10mm de largura e espessura variando de 3mm no centro a 5mm nas extremidades, com seis orifícios. A placa do compósito foi empregada na fixação de osteotomia de fêmur em quatro gatos, totalizando seis intervenções. Verificou-se a ruptura de cinco placas (83,3 por cento) até o quarto dia e de uma placa (16,7 por cento) aos 21 dias, quando se observou um calo ósseo exuberante. O resultado da implantação da placa no gato mostrou que o compósito não possui resistência suficiente para ser empregado como placas de fixação de fêmur em gatos.

The composite of polyhydroxybutyrate (PHB) 70 percent and hydroxyapatite (HA) 30 percent was evaluated as plate for bone fixation in cats. The employed composite plates presented six orifices and measured 60 x 10mm, length and width, respectively, with thickness ranging from 3 to 5mm according to the region. The composite plate was used in the fixation of femoral osteotomy in four cats, in a total of six interventions. There were ruptures in five plates (83.3 percent) until day 4 and in one plate (16.7 percent) on the day 21, when it was possible to observe an exuberant osseous callus. The result of the plate deployment in the cat showed that the composite does not have sufficient strength to be used as plate of femoral fixation in cats.

Production of PHB (polyhydroxybutyrate) by rhodopseudomonas palustris KU003 under nitrogen limitation.

A survey of various tannery effluents for the presence of purple non-sulphur bacteria was undertaken in Warangal district of South India. In all the nine bacterial species, which included Rhodopseudomonas palustris, R.rutila, R.acdiophila, Rhodopila globiformis, Rhodospirillum rubrum, Rsp.photometricum, Rhodobacter sphaeroides, Rb.capsulatus, Rhodobacter sp and Rhodocyclus gelatinosus were isolated. Among these Rhodopseudomonas palustris KU003 was selected for the production of Polyhydroxybutyrate (PHB). Effect of nitrogen limitation on the production of PHB was tested. PHB accumulation was more at a nitrogen limitation of 78 mg/L of ammonium chloride. The maximum PHB produced was 180 mg/L of BP medium containing glucose as carbon source. Significance of the above in the light of existing literature is discussed in this communication.

Biocompatibility, osteointegration, osteoconduction, and biodegradation of a hydroxyapatite-polyhydroxybutyrate composite

In this work, biocompatibility, osteointegration, osteoconductivity, and biodegradation of a hydroxyapatite polyhydroxybutyrate new composite were evaluated. The composite was implanted in rabbits' bone defects and clinical, radiographic, histological, and histomorphometric data of these animals were compared with those of unfilled defects on the days 8th, 45th, and 90th after surgery. No significant differences existed between the groups for the evaluated clinical parameters. Radiographs showed bone-composite direct contact. Bone formed within the defect, interface and inside the composite. Significant differences were found between the bone and connective tissues percentage within the defect at all dates and at the interface on the 45th day, bone tissue prevailing. Composite's biodegradation signs were evident: giant cells on the surface of composite fragments separated from the original block in the absence of inflammatory infiltrate. These data supported that such composite was biocompatible, biodegradable, osteoconductive and integrate to bone.

A biocompatibilidade, osteointegração, osteocondução e biodegradação de um novo compósito de hidroxiapatita e polihidroxibutirato foram avaliados. O compósito foi implantado em defeitos ósseos em coelhos e dados clínicos, radiográficos, histológicos e histomorfométricos foram comparados aos de defeitos não preenchidos aos 8, 45 e 90 dias após a cirurgia. Não foram observadas diferenças significantes entre os grupos para os parâmetros clínicos avaliados. Contato direto entre osso e compósito foi observado nas radiografias. Tecido ósseo se formou dentro do defeito, interface e dentro do compósito. Foram observadas diferenças significativas entre a porcentagem dos tecidos ósseo e conjuntivo dentro do defeito em todas as datas de avaliação e na interface aos 45 dias, com predominância do tecido ósseo. Sinais de biodegradação foram observados: células gigantes na superfície de fragmentos do compósito separados do bloco original, na ausência de infiltrado inflamatório. Os dados permitem concluir que o compósito é biocompatível, biodegradável, osteocondutor e se integra ao tecido ósseo.

Compósito para fabricação de implantes reabsorvíveis para osteossíntese: avaliação da biocompatibilidade em coelhos / Composite for the fabrication of resorbable implants for osteosynthesis: biocompatibility evaluation in rabbits

A biocompatibilidade, a osseointegração e a reabsorção de compósitos contendo 10 por cento de hidroxiapatita (HAP-91®) e 90 por cento de polihidroxibutirato foram analisadas após implantação em defeitos ósseos em coelhos. Foram obtidos dados clínicos, radiográficos, histológicos e histomorfométricos aos oito, 45 e 90 dias após a realização da cirurgia, sendo os resultados comparados aos de defeitos não-preenchidos. Não foram observadas diferenças significativas entre os grupos para os parâmetros clínicos avaliados. Radiografias demonstraram o compósito em contato direto com osso aos 45 e 90 dias. Tecido ósseo se formou normalmente no defeito e na interface com o material, quando comparado ao controle. Não foi observado infiltrado inflamatório. Em todos os dias, a porcentagem de tecido ósseo foi maior que a de tecido conjuntivo (P=0,001 aos 8 e 45 dias e 0,022 aos 90 dias) dentro dos defeitos. Foram observados tecidos ósseo e conjuntivo na interface com o compósito, sendo o último significativamente maior que o primeiro aos 90 dias (P=0,004). Foram observados sinais de reabsorção do compósito: células gigantes na superfície de fragmentos do compósito separados do bloco original. Portanto, pode-se concluir que o compósito é biocompatível em coelhos, reabsorvível e se integra ao tecido ósseo.

Biocompatibility, osseointegration, and resorption of a 10 percent hydroxyapatite (HAP-91®) - 90 percent polyhydroxybutyrate composite were analyzed after implantation in rabbits´ bone defects. Clinical, radiographic, histologic and histomorphometric data were collected on the 8th, 45th, and 90th days after surgery and the results were compared to those of unfilled defects. No significant differences between groups for the evaluated clinical parameters existed. Radiographs showed bone-composite direct contact. Bone tissue was formed normally in the defect and at the composite-tissue interface when compared to the control. No inflammatory infiltrated was observed. For each sampling day, the percentages of bone were greater than those of connective tissues (P=0.001 on the 8th and 45th days and 0.022 on the 90th day) inside the defect. Bone and connective tissues were seen at the interface with the composite, being the last one significantly greater than the first on the 90th day (P=0.004). Composite's resorption signs were evident: giant cells on the surface of composite fragments separated from the original bloc. Data support that such composite is biocompatible in rabbits, resorbable and integrate to bone.

Polyhydroxybutyrate production using agro-industrial residue as substrate by Bacillus sphaericus NCIM 5149

The aim of this work was to study the production of polyhydroxybutyrate (PHB) using agro- industrial residues as the carbon source. Seven substrates, viz., wheat bran, potato starch, sesame oil cake, groundnut oil cake, cassava powder, jackfruit seed powder and corn flour were hydrolyzed using commercial enzymes and the hydrolyzates assessed for selecting the best substrate for PHB production. Jackfruit seed powder gave the maximum production of PHB under submerged fermentation using Bacillus sphaericus (19 percent) at the initial pH of 7.5.

System of Intein-mediated PHB Purified Human Antimicrobial Peptide LL-37 / 中国生物工程杂志

The new intein-mediated PHB purify protein system is a high expression, automatic cutting, for purification, low-cost protein purification system,it is conducive to large-scale protein purification.Choose human antibacterial peptide LL-37 as the purification objects,which is poison to prokaryotic cell.We construct intein-mediated PHB purified human antimicrobial peptide LL-37 system through genetic engineering technology and use this system to purify LL-37. The results show that this system can highly express LL-37 fusion protein and purifiy the product as same size with expectations.