Characteristics and functional outcomes of varus displaced proximal humerus fractures.

BACKGROUND: The purpose of this study was to compare fracture characteristics and functional outcomes between patients with proximal humerus fractures with and without initial varus displacement. METHODS: A retrospective review of 325 patients with proximal humerus fractures was performed. Patients with initial varus displacement were placed in Varus cohort and were age- and sex-matched 1:1 with a second cohort presenting proximal humerus fractures without varus displacement, referred to as Fracture cohort. Varus fracture displacement was defined when the most proximal aspect of humeral head was below the most proximal aspect of greater tuberosity on initial radiographs, and the head shaft angle was <130°. RESULTS: There were 60 patients in V cohort and 60 patients in F cohort. Statistical analysis revealed that there were significant differences in initial horizontal offset (38.8 vs. 45.9 mm), initial anterior angulation angle (36.5° vs. 16.4°), postoperative head shaft angle (132.2° vs. 141.3°), last head shaft angle (122.2° vs. 138.5°), difference for head shaft angles (10.0° vs. 2.7°), postoperative horizontal offset (43.4 vs. 45.3 mm), last horizontal offset (38.4 vs. 42.8 mm), difference for offsets (4.9 vs. 2.5 mm), complications (15 vs. 7 cases), and revision surgery (7 vs. 1 case) between two cohorts. Overall satisfactory results were achieved in most patients regardless of varus displacement, pain-VAS and Constant scores in V cohort were inferior to the scores in F cohort. The cut-off value of postoperative head shaft angle for good/excellent outcomes was 135.5° using receiver operating characteristic curve analyses. CONCLUSION: Varus displaced proximal humerus fractures were accompanied by decreased horizontal offset and increased anterior angulation angle, and had a course of more varization and horizontal shortening compared with those without initial varus displacement. Patients with varus displaced fractures were associated with worse functional outcomes, and these factors might affect functional outcomes. LEVEL OF EVIDENCE: Prognostic, cohort study, Level III.

Surgical outcomes of segmental diaphyseal forearm fractures in adults: a small case series on plate osteosynthesis, intramedullary nailing, and other surgical methods.

BACKGROUND: Segmental fractures often result from high-energy or indirect trauma that causes bending or torsional forces with axial loading. We evaluated surgical outcomes of patients with forearm segmental diaphyseal fractures. METHODS: We retrospectively analyzed data from patients with forearm segmental fractures for which they underwent surgery at the Pusan National University Trauma Center from March 2013 to March 2022. We also analyzed accompanying injuries, injury severity score (ISS), injury mechanism, occurrence of open fracture, surgical technique, and treatment results. RESULTS: Fifteen patients were identified, one with bilateral segmental diaphyseal forearm bone fracture, for a total of 16 cases. Nine of the patients were male. The overall mean age was 50 years, and the mean follow-up period was 16.2 months. Six cases who underwent surgery using plate osteosynthesis achieved bone union without length deformity at final follow-up. Three of seven patients who underwent intramedullary nailing alone underwent reoperation due to nonunion. Six cases achieved bone union at final follow-up, three of which showed length deformity. Three patients underwent surgery using a hybrid method of IM nailing, plates, and mini cables. One patient who underwent surgery with a plate and one patient who underwent surgery with IM nailing alone showed nonunion and were lost to follow-up. CONCLUSION: Plate osteosynthesis is considered the gold standard for treatment of adult forearm diaphyseal segmental fractures. In this study, IM nailing was associated with high rates of non-union and length deformity. However, the combination of IM nailing and a plate-cable system may be an acceptable alternative in segmental diaphyseal forearm fracture, achieving a union rate similar to that provided by plate fixation.

Usefulness of Percutaneous Pedicle Screw Fixation for Treatment of Lower Lumbar Burst (A3-A4) Fractures: Comparative Study with Thoracolumbar Junction Fractures.

Background: Percutaneous pedicle screw fixation (PPSF) without fusion has been recently recommended in the treatment of thoracolumbar fracture to reduce the adverse effects associated with the conventional open approaches and to restore range of motion. However, those studies report on the thoracolumbar junction, and there is no report on lower lumbar fracture. Purpose: To assess effectiveness of PPSF without fusion for treating lower lumbar burst (A3 and A4) fractures. Methods: A retrospective analysis was made to evaluate consecutive 50 patients with AO type A3 and A4 thoracolumbar fracture underwent PPSF. Patients were divided into a thoracolumbar junction (TLJ) group (T11-L2) and lower lumbar (LL) group (L3-5). The following items were measured and compared between the two groups. Vertebral height and consolidation, retropulsed fragment, sagittal curve and fixation failure were assessed with certain interval regularly. Results: The average height at pre- and post-reduction were 56.2% (36.2-74.3), 95.3% (84.2-98.3) in TLJ group and 65.7% (45.7-86.2), 91% (73.1-100) in LL group. The average canal area occupancy rate at pre- and post-reduction were 46.1% (37.4%-67.5%), 38.1% (31.3%-40.8%) in TLJ group and 40.4% (15.0-65.7), 19.3% (9.4-26.6) in LL group. Consolidation was completed within 12 months after surgery in both groups. There was no significant difference between two groups in clinical and radiographic parameters except cobb angle loss. Conclusion: Patients with lower lumbar fracture can be effectively managed with PPSF without fusion. PPSF following the implant removal can restore the movement of the lower lumbar spine, which is essential for daily life.

Polyhydroxybutyrate (PHB) production from sugar cane molasses and tap water without sterilization using novel strain, Priestia sp. YH4.

The production cost of biodegradable polymer like polyhydroxybutyrate (PHB) is still higher than that of petroleum-based plastics. A potential solution for reducing its production cost is using a cheap carbon source and avoiding a process of sterilization. In this study, a novel PHB-producing microbial strain, Priestia sp. YH4 was screened from the marine environment using sugarcane molasses as the carbon source without sterilization. Culture conditions, such as carbon, NaCl, temperature, pH, inoculum size, and cultivation time, were optimized for obtaining the highest PHB production by YH4 resulting in 5.94 g/L of dry cell weight (DCW) and 61.7 % of PHB content in the 5 mL culture. In addition, it showed similar PHB production between the cultures with or without sterilization in Marine Broth media. When cultured using only tap water, sugarcane molasses, and NaCl in a 5 L fermenter, 24.8 g/L DCW was produced at 41 h yielding 13.9 g/L PHB. Finally, DSC (Differential Scanning Calorimetry) and GPC (Gel Permeation Chromatography) were used to analyze thermal properties and molecular weights resulting in Tm = 167.2 °C, Tc = 67.3 °C, Mw = 2.85 × 105, Mn = 1.05 × 105, and PDI = 2.7, respectively. Therefore, we showed the feasibility of more economical process for PHB production by finding novel strain, utilizing molasses with minimal media components and avoiding sterilization.

Wide Resection Preserving Conjoined Tendons of the Rotator Cuff Muscles for Undifferentiated Pleomorphic Sarcoma at the Suprascapular Fossa - A Case Report.

Surgical principles in sarcoma are a wide resection, including surrounding tissues and maximisation of the function of the affected limb. Rotator cuff muscles are biomechanically important structures acting as a force couple in movement of the shoulder joint. Thus, conjoined tendons are essential for motion capability in absence of the supraspinatus muscle. This article reports a case of a large undifferentiated pleomorphic sarcoma (UPS) at the suprascapular fossa in a 78-year-old man. After diagnosis of sarcoma, he underwent wide, en-bloc excision preserving conjoined tendons of rotator cuff muscles and low-dose radiation therapy for surveillance of local recurrence. All dissection was performed to avoid contaminating the tumour and involved the whole supraspinatus except the conjoined tendons. We report a case of UPS at the suprascapular fossa, which showed a good result after a wide resection preserving conjoined tendons of rotator cuff muscles. Level of Evidence: Level V (Therapeutic).

Risk factors for nonunion in oblique lateral interbody fusion.

BACKGROUND: Compared with posterior interbody fusion techniques, oblique lateral interbody fusion (OLIF) offers a larger fusion bed with greater intervertebral space access, use of larger cages, more sufficient discectomy, and better end-plate preparation. However, the fusion rate of OLIF is similar to that of other interbody fusions. This study aimed to examine the factors associated with nonunion in OLIF. METHODS: This study examined 201 disc levels from 124 consecutive patients who underwent OLIF for lumbar degenerative diseases with 1-year regular follow-up. Demographic and surgical factors were reviewed from the medical records. Radiological factors measured were sagittal parameters, intervertebral disc angle (DA) before surgery and at the final follow-up, presence of vertebral end-plate lesions, and cage subsidence. Multivariable logistic regression analysis was performed to identify the factors associated with nonunion. RESULTS: Among the 201 discs, 185 (92.0%) achieved union at 1-year followed up. Smoking, surgery at the L5-S1 level, not performing laminectomy, and a large intervertebral DA were factors associated with nonunion in OLIF (all P < 0.05). Multivariable logistic regression analysis showed two independent variables (surgery at L5-S1 level and not performing laminectomy) as risk factors for nonunion in OLIF. CONCLUSIONS: Not performing laminectomy and surgery at the L5-S1 level were risk factors for nonunion in OLIF. To reduce the nonunion rate, surgeons should consider additional stabilization strategies for the L5-S1 OLIF and perform laminectomy.

Enhanced tolerance of Cupriavidus necator NCIMB 11599 to lignocellulosic derived inhibitors by inserting NAD salvage pathway genes.

Polyhydroxybutyrate (PHB) is a bio-based, biodegradable and biocompatible plastic that has the potential to replace petroleum-based plastics. Lignocellulosic biomass is a promising feedstock for industrial fermentation to produce bioproducts such as polyhydroxybutyrate (PHB). However, the pretreatment processes of lignocellulosic biomass lead to the generation of toxic byproducts, such as furfural, 5-HMF, vanillin, and acetate, which affect microbial growth and productivity. In this study, to reduce furfural toxicity during PHB production from lignocellulosic hydrolysates, we genetically engineered Cupriavidus necator NCIMB 11599, by inserting the nicotine amide salvage pathway genes pncB and nadE to increase the NAD(P)H pool. We found that the expression of pncB was the most effective in improving tolerance to inhibitors, cell growth, PHB production and sugar consumption rate. In addition, the engineered strain harboring pncB showed higher PHB production using lignocellulosic hydrolysates than the wild-type strain. Therefore, the application of NAD salvage pathway genes improves the tolerance of Cupriavidus necator to lignocellulosic-derived inhibitors and should be used to optimize PHB production.

A Modified Anterior Column Realignment With Partial Anterior Longitudinal Ligament Release in Oblique Lateral Interbody Fusion.

STUDY DESIGN: Retrospective radiological analysis. OBJECTIVE: To demonstrate the radiological outcome after a modified anterior column realignment (mACR) with partial anterior longitudinal ligament (ALL) release in oblique lateral interbody fusion (OLIF). SUMMARY OF BACKGROUND DATA: Anterior column realignment (ACR) remains a powerful sagittal correction technique in minimally invasive adult spinal deformity surgery and is often combined with posterior column osteotomy (PCO) to achieve more lordosis. OLIF is ideal for ACR because the anterior-to-psoas corridor typically involves the anterolateral half of the disk. METHODS: This study included 112 operated disk levels of 101 consecutive patients who underwent OLIF between L2-L3 and L4-L5 using a 12° lateral cage. The mACR was performed at 73 (65.2%) levels with 30% to 50% sectioning of the ALL. Each operated level was grouped according to the mACR and additional PCO as: (1) no mACR, OLIF only (n=39); (2) mACR with no PCO (n=18); (3) mACR with grade 1 PCO (n=27); (4) mACR with grade 2 PCO (n=22); or (5) mACR with grade 3 PCO (n=6). RESULTS: At the last follow-up, the mean disk lordotic angles were 10.9±2.9°, 12.6±3.0°, 13.3±3.9°, 16.7±3.2°, and 16.8±2.4° in the no mACR, mACR with no PCO, mACR with grade 1 PCO, mACR with grade 2 PCO, and mACR with grade 3 PCO groups, respectively ( P <0.001). The mean increases in disk lordotic angle were 5.8±4.1°, 12.1±6.1°, 13.5±8.7°, 15.8±6.7°, and 17.9±6.2° in each group, respectively ( P <0.001). CONCLUSIONS: ACR can be performed with partial ALL release under direct vision in OLIF without deep dissection into the ventral disk space. The mACR in OLIF is a simple, safe, and effective technique for anterior column lengthening. LEVEL OF EVIDENCE: 4.

Bioconversion of Mixed Alkanes to Polyhydroxyalkanoate by Pseudomonas resinovornas: Upcycling of Pyrolysis Oil from Waste-Plastic.

Polyhydroxyalkanoate (PHA) is a biodegradable plastic that can be used to replace petroleum-based plastic. In addition, as a medium-chain-length PHA (mcl-PHA), it can be used to provide elastomeric properties in specific applications. Because of these characteristics, recently, there has been much research on mcl-PHA production using inexpensive biomass materials as substrates. In this study, mcl-PHA producers were screened using alkanes (n-octane, n-decane, and n-dodecane) as sources of carbon. The amount of PHA produced by Pseudomonas resinovorans using sole n-octane, n-decane, or n-dodecane was 0.48 g/L, 0.27 g/L, or 0.07 g/L, respectively, while that produced using mixed alkane was 0.74 g/L. As a larger amount of PHA was produced using mixed alkane compared with sole alkane, a statistical mixture analysis was used to determine the optimal ratio of alkanes in the mixture. The optimal ratio predicted by the analysis was a medium with 9.15% n-octane, 6.44% n-decane, and 4.29% n-dodecane. In addition, through several concentration-specific experiments, the optimum concentrations of nitrogen and phosphorus for cell growth and maximum PHA production were determined as 0.05% and 1.0%, respectively. Finally, under the determined optimal conditions, 2.1 g/L of mcl-PHA and 60% PHA content were obtained using P. resinovorans in a 7 L fermenter.

Humeral head coverage in arthroscopic partial repair of massive rotator cuff tears improves functional outcomes: an analysis of influential factors.

BACKGROUND: Although partial rotator cuff repair has shown good outcomes, differences in clinical outcomes remain concerns. This study was performed to determine whether patients with humeral head coverage would show better functional outcomes than patients without humeral head coverage and to identify the factors for humeral head coverage after arthroscopic partial repair of massive tears. METHODS: We performed a retrospective study of 63 patients with massive rotator cuff tears who underwent arthroscopic partial repair between 2012 and 2018. Two to four margin convergences were first performed; then, the Mason-Allen technique was performed. The patients were divided into 2 groups: those with humeral head coverage (38 cases) and those without humeral head coverage (25 cases). The following factors were evaluated: age; sex; hypertension; diabetes; osteoporosis; preoperative and postoperative pseudoparalysis, visual analog scale (VAS) pain score, Constant score, acromiohumeral distance, and subacromial bony spur; and subscapularis tear and repair. Muscle atrophy and fatty degeneration were evaluated by magnetic resonance imaging preoperatively, and the integrity of the repaired cuff was evaluated by ultrasonography at a minimum of 2 years after surgery. RESULTS: Compared with preoperative values, significant improvements in VAS pain scores (from 6.27 to 2.32 in patients with humeral head coverage and from 7.00 to 2.81 in those without humeral head coverage) and Constant scores (from 51.35 to 75.95 and from 44.62 to 69.81, respectively) were observed in both groups (P < .001). Statistical analysis revealed that postoperative VAS pain scores (2.32 vs. 2.81) and Constant scores (75.95 vs. 69.81) in patients with humeral head coverage were superior to those in patients without humeral head coverage (P = .044 and P = .003, respectively). The integrity of the repaired cuff was evaluated by ultrasonography, and partial tears were found in 4 of 37 patients with humeral head coverage and 2 of 26 patients without humeral head coverage (P = .816). Univariable logistic regression analysis revealed that age (P < .001), comorbidity (P = .005), symptom duration (P = .023), preoperative shoulder mobility (P < .001), maintained acromiohumeral distance (P = .006), subscapularis tear (P = .026), and less preoperative supraspinatus and infraspinatus muscle atrophy (P = .001 and P = .010, respectively) had significant correlations with humeral head coverage. CONCLUSIONS: Overall satisfactory results were achieved in most patients regardless of high retear rates, but patients with partial repair covering the humeral head were associated with better outcomes than patients without humeral head coverage. Multivariable regression analysis revealed that age (<70 years, P = .003), capability of shoulder mobility (P = .005), maintenance of the acromiohumeral space (>7 mm, P = .016), and less atrophy of the rotator cuff muscles (P = .021) were favorable factors to achieve humeral head coverage during surgical partial repair of massive rotator cuff tears.

Development of novel on-line capillary gas chromatography-based analysis method for volatile organic compounds produced by aerobic fermentation.

Many volatile compounds, such as isoprene, a precursor used in the synthesis of natural rubber, have been produced through fermentation using genetically engineered microorganisms. Despite this biotechnological success, measuring the concentrations of volatile compounds during fermentation is difficult because of their high volatility. In current systems, off-line analytical methods usually lead to product loss, whereas on-line methods raise the production cost due to the requirement of complex devices. Here, we developed a novel on-line gas chromatography (GC)-based system for analyzing the concentration of isoprene with the aim to minimize the cost and requirement for devices as compared to current strategies. In this system, a programmable logic controller is used to combine conventional GC with a syringe pump module (SPM) directly connected to the exhaust pipe of the fermentor, and isoprene-containing samples are continuously pumped from the SPM into the GC using an air cylinder recycle stream. We showed that this novel system enables isoprene analysis during fermentation with convenient equipment and without the requirement of an expensive desorption tube. Furthermore, this system may be extended to the detection of other volatile organic compounds in fermentation or chemical processes.

Production of (3-hydroxybutyrate-co-3-hydroxyhexanoate) copolymer from coffee waste oil using engineered Ralstonia eutropha.

Polyhydroxyalkonate (PHA) is a type of polymer that has the potential to replace petro-based plastics. To make PHA production more economically feasible, there is a need to find a new carbon source and engineer microbes to produce a commercially valuable polymer. Coffee waste is an inexpensive raw material that contains fatty acids. It can act as a sustainable carbon source and seems quite promising with PHA production in Ralstonia eutropha, which is a well-known microbe for PHA accumulation, and has the potential to utilize fatty acids. In this study, to make poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(HB-co-HHx)), which has superior properties in terms of biodegradability, biocompatibility, and mechanical strength, engineered strain Ralstonia eutropha Re2133 overexpressing (R)-specific enoyl coenzyme-A hydratase (phaJ) and PHA synthetase (phaC2) with deletion of acetoacetyl Co-A reductases (phaB1, phaB2, and phaB3) was used to produce PHA from coffee waste oil. At a coffee oil concentration of 1.5%, and C/N ratio of 20, the R. eutropha Re2133 fermentation process results in 69% w/w of DCW PHA accumulation and consists of HB (78 mol%) and HHx (22 mol%). This shows the feasibility of using coffee waste oil for P(HB-co-HHx) production, as it is a low-cost fatty acid enriched waste material.

A Hierarchically Modified Graphite Cathode with Au Nanoislands, Cysteamine, and Au Nanocolloids for Increased Electricity-Assisted Production of Isobutanol by Engineered Shewanella oneidensis MR-1.

It is necessary to understand the surface structural effects of electrodes on the bioalcohol productivity of Shewanella oneidensis MR-1, but this research area has not been deeply explored. Here, we report that the electricity-assisted isobutanol productivity of Shewanella oneidensis MR-1::pJL23 can be enhanced by sequentially modifying a graphite felt (GF) surface with Au nanoislands (Au), cysteamine (NH2), and Au nanoparticles (Au NPs). After bacteria were incubated for 50 h with the unmodified GF under various electrode potentials (vs Ag/AgCl), the bacterial isobutanol concentrations increased from 2.9 ± 1 mg/L under no electricity supply to a maximum of 5.9 ± 1 mg/L at -0.6 V. At this optimum electrode potential, the concentrations continued increasing to 9.1 ± 1, 14 ± 2, and 27 ± 2 mg/L when the GF electrodes were modified with Au, NH2-Au, and Au NP-NH2-Au, respectively. We further studied how each surface structure affected the bacterial adsorptions, current profiles, and biofilms' electrochemical performances. In particular, these modifications induced the adsorption of elongated bacteria, with the amount dependent on the electrode structure. In the presence of electric supply, the amount of elongated bacteria further increased. We also found that the NH2-Au-GF and Au NP-NH2-Au-GF electrodes themselves could increase the concentrations to 11 ± 0.3 and 12 ± 2 mg/L, respectively, upon the bacterial incubation without electricity. Among the electrodes tested, the contribution of electricity to the bacterial isobutanol production was the greatest with the Au NP-NH2-Au-GF electrode. After 96 h of incubation, the concentration increased to 72 ± 2 mg/L, which was 4.7 and 3.7 times the previously reported values obtained without and with electricity, respectively.

Microbial biodiesel production from oil palm biomass hydrolysate using marine Rhodococcus sp. YHY01.

The effect of various biomass derived inhibitors (i.e. furfural, hydroxymethylfurfural (HMF), vanillin, 4-hydroxy benzaldehyde (4-HB) and acetate) was investigated for fatty acid accumulation in Rhodococcus sp. YHY 01. Rhodococcus sp. YHY01 was able to utilize acetate, vanillin, and 4-HB for biomass production and fatty acid accumulation. The IC50 value for furfural (3.1mM), HMF (3.2mM), vanillin (2.0mM), 4-HB (2.7mM) and acetate (3.7mM) was calculated. HMF and vanillin affect fatty acid composition and increase saturated fatty acid content. Rhodococcus sp. YHY 01 cultured with empty fruit bunch hydrolysate (EFBH) as the main carbon source resulted in enhanced biomass (20%) and fatty acid productivity (37%), in compression to glucose as a carbon source. Overall, this study showed the beneficial effects of inhibitory molecules on growth and fatty acid production, and support the idea of biomass hydrolysate utilization for biodiesel production by avoiding complex efforts to remove inhibitory compounds.

Medium engineering for enhanced production of undecylprodigiosin antibiotic in Streptomyces coelicolor using oil palm biomass hydrolysate as a carbon source.

In this study, a biosugar obtained from empty fruit bunch (EFB) of oil palm by hot water treatment and subsequent enzymatic saccharification was used for undecylprodigiosin production, using Streptomyces coelicolor. Furfural is a major inhibitor present in EFB hydrolysate (EFBH), having a minimum inhibitory concentration (MIC) of 1.9mM, and it reduces utilization of glucose (27%), xylose (59%), inhibits mycelium formation, and affects antibiotic production. Interestingly, furfural was found to be a good activator of undecylprodigiosin production in S. coelicolor, which enhanced undecylprodigiosin production by up to 52%. Optimization by mixture analysis resulted in a synthetic medium containing glucose:furfural:ACN:DMSO (1%, 2mM, 0.2% and 0.3%, respectively). Finally, S. coelicolor was cultured in a fermenter in minimal medium with EFBH as a carbon source and addition of the components described above. This yielded 4.2µg/mgdcw undecylprodigiosin, which was 3.2-fold higher compared to that in un-optimized medium.

Production of rapamycin in Streptomyces hygroscopicus from glycerol-based media optimized by systemic methodology.

Rapamycin, produced by the soil bacterium Streptomyces hygroscopicus, has the ability to suppress the immune system and is used as an antifungal, anti-inflammatory, antitumor, and immunosuppressive agent. In an attempt to increase the productivity of rapamycin, mutagenesis of wild-type Streptomyces hygroscopicus was performed using ultraviolet radiation, and the medium composition was optimized using glycerol (which is one of the cheapest starting substrates) by applying Plackett-Burman design and response surface methodology. Plackett-Burman design was used to analyze 14 medium constituents: M100 (maltodextrin), glycerol, soybean meal, soytone, yeast extract, (NH4)2SO4, L-lysine, KH2PO4, K2HPO4, NaCl, FeSO4·7H2O, CaCO3, 2-(N-morpholino) ethanesulfonic acid, and the initial pH level. Glycerol, soytone, yeast extract, and CaCO3 were analyzed to evaluate their effect on rapamycin production. The individual and interaction effects of the four selected variables were determined by Box-Behnken design, suggesting CaCO3, soytone, and yeast extract have negative effects, but glycerol was a positive factor to determine rapamycin productivity. Medium optimization using statistical design resulted in a 45% (220.7 ± 5.7 mg/l) increase in rapamycin production for the Streptomyces hygroscopicus mutant, compared with the unoptimized production medium (151.9 ± 22.6 mg/l), and nearly 588% compared with wildtype Streptomyces hygroscopicus (37.5 ± 2.8 mg/l). The change in pH showed that CaCO3 is a critical and negative factor for rapamycin production.

Phosphorylation of chloramphenicol by a recombinant protein Yhr2 from Streptomyces avermitilis MA4680.

Although phosphorylation of chloramphenicol has been shown to occur in the chloramphenicol producer, Streptomyces venezuelae, there are no reports on the existence of chloramphenicol phosphorylase in other Streptomyces species. In the present study, we report the modification of chloramphenicol by a recombinant protein, designated as Yhr2 (encoded by SAV_877), from Streptomyces avermitilis MA4680. Recombinant Yhr2 was expressed in Escherichia coli BL21 (DE3) and the cells expressing this recombinant protein were shown to phosphorylate chloramphenicol to a 3'-O-phosphoryl ester derivative, resulting in an inactivated form of the antibiotic. Expression of yhr2 conferred chloramphenicol resistance to E. coli cells up to 25 µg/mL and in an in vitro reaction, adenosine triphosphate (ATP), guanosine triphosphate (GTP), adenosine diphosphate (ADP) and guanosine diphosphate (GDP) were shown to be the phosphate donors for phosphorylation of chloramphenicol. This study highlights that antibiotic resistance conferring genes could be easily expressed and functionalized in other organisms that do not produce the respective antibiotic.

Identification and functional characterization of an α-amylase with broad temperature and pH stability from Paenibacillus sp.

Amylases are important industrial enzymes that have been applied widely in the food, detergent, and pulp industries and fermentation processes. In the present study, a gene encoding an alpha-amylase from the genomic DNA library of Paenibacillus sp. was identified and characterized. The amylase gene designated amy1 was shown to consist of 1,980 bp and shared sequence identity towards α-amylase genes from other Bacillus sp. The deduced amino acid sequence for Amy1 indicated 80 % sequence identity with other Bacillus strains. Heterologous expression of recombinant Amy1 in Escherichia coli BL21(DE3) facilitated the recovery of this protein in soluble form. Enzyme kinetic data revealed Amy1 to have a K m of 23.83 mg/mL and K cat of 48.74 min(-1) and K cat /K m of 2 min(-1) mg(-1) mL(-1) for starch. The activity of this protein was found to be enhanced by Mn(2+), and furthermore, Amy1 remained active at a broad pH range (4-10) and temperature (30-90 °C). The ability of Amy1 to act on food waste under broad temperature and pH conditions, together with its ability to produce simple sugars, shows many advantages for further application in the food industry.