We tested the ability of 14 strains of Trichoderma to emit volatile compounds that decreased or stopped the growth of Phytophthora infestans. Volatile organic compounds (VOCs) emitted from Trichoderma strains designated T41 and T45 inhibited the mycelial growth of P. infestans grown on a laboratory medium by 80 and 81.4%, respectively, and on potato tubers by 93.1 and 94.1%, respectively. Using the DNA sequence analysis of the translation elongation factor region, both Trichoderma strains were identified as Trichoderma atroviride. VOCs emitted by the strains were analyzed, and 39 compounds were identified. The most abundant compounds were 3-methyl-1-butanol, 6-pentyl-2-pyrone, 2-methyl-1-propanol, and acetoin. Electron microscopy of the hyphae treated with T. atroviride VOCs revealed serious morphological and ultrastructural damages, including cell deformation, collapse, and degradation of cytoplasmic organelles. To our knowledge, this is the first report describing the ability of Trichoderma VOCs to suppress the growth of the late blight potato pathogen.

BackgroundAlpha-amylases hydrolyze 1,4 α-glycosidic bonds of starch and produce malto-oligosaccharides. It is an important enzyme generally applied in textile, food and brewing industries. Enhancement in thermal stability and productivity of enzymes are the two most sought after properties for industrial use. The Aspergillus oryzae (Koji) has Generally Recognized as Safe (GRAS) status and safe for use in food industry. Hence, Koji strain’s development for the screening of potent mutants, hyper producer of thermostable α-amylases, with desired attributes is the need of the time.ResultsA process has been developed to improve super Koji (A. oryzae cmc1) strain through γ-rays treatment. The doses i.e. 0.60, 0.80, 1.00, 1.20 & 1.40 KGy gave more than 3.0 log kill. Initially, 52 Koji mutants resistant to 1% (w/v) Triton X-100 were selected. 2nd screening was based on α-amylases hyper production and 23 mutants were sorted out by measuring clearing zones index (CI). Afterwards nine potent mutants, resistant to 2-deoxy D-glucose, were screened based on CI. These were further analyzed for thermal stability and productivity of α-amylase under submerged conditions. The mutants’ M-80(10), M-100(6) & M-120(5) gave about four fold increases in α-amylases productivity. The half life of M-100(6) α-amylase at 55 °C was 52 min and was highest among the mutants. Liquid Chromatography-Mass Spectrometry (LC-MS) analysis confirmed that mutants did not produce aflatoxins. Field Emission Scanning Electron Microscopy (FESEM) of Koji mycelia depicted that exposure to gamma rays increased rigidity of the mycelium. The potent Koji mutant M-100(6) was grown on soluble starch in 10L fermenter and produced 40.0 IU ml-1 of α-amylases with specific activity of 2461 IU mg-1 protein. Growth kinetic parameters were: μ = Specific growth rate= 0.069 h-1, td = Biomass doubling time= 10.0 h, Yp/x = Product yield coefficient with respect to cell mass = 482 U g-1; qp= Specific rate of product formation= 33.29 U g-1 h-1.ConclusionIt was concluded that the developed five step screening process has great potential to generate potent mutants for the hyper production of thermostable enzymes through γ-rays mediated physical mutagenesis. The developed thermostable α-amylases of super Koji mutantM-100(6) has immense potential for application in saccharification process for maltose syrup production. Moreover, the developed five step strain’s development process may be used for the simultaneous improvement in productivity and thermal stability of other microbial enzymes.Electronic supplementary materialThe online version of this article (10.1186/s12866-018-1345-y) contains supplementary material, which is available to authorized users.

No abstract

Background The β-1, 4-endoglucanase belongs to cellulases group of enzymes, which hydrolyse β-1, 4 glucosidic bonds in the naturally occurring cellulose or synthetic Carboxymethyl cellulose CMC thus also named as CMCase. Endoglucanase from Aspergillus oryzae is favoured in food and feed industries due to its GRAS (generally regarded as safe) status and for relatively stable enzymes production. Results Current report is novel as it deals with the development of super Koji strain having hyper production of thermostable CMCases. Hence, the potent Koji’s mutant, hyper producer of thermostable endoglucanase was screened from the available mutants stock (Total = 52 variants), which was generated through gamma rays treatment of Super Koji strain (A. oryzae cmc1). Initially, seventeen (17) mutants hyper producer of CMCases were selected based on the basis of clearance zone index (CI) on the plates supplemented with 1% CMC. Afterwards, six mutants resistant to 2-deoxy-D-glucose were selected having enhanced CMCase production. Mutants’ next screening was on submerged CMCase production. Potent mutant 60(5) having 60 kRad γ-rays exposure showed 3.6 folds hyper CMCase production (6.02 U ml-1). The half-life of endoglucanse produce of A. oryzae mutant 60(5) was 29.42 min and 14.8 min at 55 ⁰C and 60 ⁰C, respectively, which was 4 folds higher than the control at 60 ⁰C. The LC-MS analysis showed that A. oryzae mutant 60(5) does not produce aflatoxins. Field Emission Scanning Electron Microscopy (FESM) of mutated Koji depicted that gamma radiations enhanced the stiffness of fungal mycelia. Kinetics of CMCase production by mutant 60(5) in 10 L and 200L fermenter was as: CMCase = 913 U dl-1 & 821 U dl-1; Specific growth (µ) = 0.064 & 0.082 hours; Cell mass doubling time (td) = 10.71 & 8.42 hours; Yield coefficient with respect to cell mass (Yp/x) = 59 Ug-1 & 89 U g-1 and Specific rate of CMCase production (qp) = 3.84 Ug-1h-1 & 7.37 Ug-1h-1 respectively. Conclusion We concluded, γ-rays induced potent mutations in mutant 60(5) resulting in hyper production of thermostable endoglucanase, which can withstand harsh industrial conditions. Hence, has great potential for applications in food & feed and other industrial processes.

e24017 Background: Lung cancer is frequently diagnosed over the age of 70 years, in a population with increased risk factors for frailty. Frailty impacts cancer treatment selection, tolerance, and quality of life. We performed a retrospective study to determine whether there is a difference in the indicators of frailty in patients of different ages, and if this impacted treatment choice. Methods: Electronic records of patients who were discussed at the University Hospitals of Leicester Lung multidisciplinary team meeting (MDT) between January to December 2019 with a suspected diagnosis of lung cancer were reviewed. Populations below the age of 65 years and over the age of 65 years were compared. Results: 846 patient records were reviewed, and 80% were aged over 65 years. Baseline WHO performance status (PS) where recorded showed 35% were PS 0-1, and 28% were 3-4. Comparatively, in the under 65 population 46% were PS 0-1, and 10% were 3-4. The over 65s had a lower rate of independence with activities of daily living (ADLs) compared to the under 65s (54% vs 71%), assistance with basic ADLs was higher (10% vs 5%) and patients with unassisted mobility was also lower (48% vs 66%). The over 65s had an increased rate of multiple comorbidities compared to the younger group (no comorbidity 25% vs 30%, 3 or more comorbidities 27% vs 14% respectively.) Where curative intent was intended, in the under 65s, surgical review deemed 24% unsuitable and of these, 72% had a discussion regarding radiotherapy with 61% receiving treatment. Comparatively in the older population, 40% of patients were deemed unsuitable for surgery and 69% went for discussion regarding radiotherapy with 44% receiving treatment. Where systemic anti-cancer therapy (SACT) discussion was advised, more patients in the under 65 group saw a medical oncologist compared to the older group (73% vs 61%) and had a higher rate of subsequent SACT (80% vs 61%). There were similar rates of non-completion of SACT in the over 65s compared to the younger group (55% vs 54%) and number of dose reductions (18% vs 15%). 35% of the over 65s and 46% of the under 65s received immunotherapy. In the older population, 30% received monotherapy, compared to 17% in the younger population. 37% of the older patients having SACT were admitted with a median stay of 2 days (range 1-30) compared to 31% in the younger group, with a median stay of 1.5 days (range 1-2). Conclusions: Our review demonstrates that in the older population, PS is higher, with a lower independence rate for ADLs, a higher level of assisted mobility and increased comorbidity. These frailty indicators impacted treatment options; less older patients were suitable for treatment compared to the younger population. The older population displayed more indicators of frailty potentially impacting treatment options, and this highlights the need for active frailty assessment and rapid intervention in the lung cancer population to allow a full range of treatment options.

In this advance era, the enzymes are considered as a core kernel of white biotechnology and their demand is increasing day by day. According to report published in Research and Markets (ID: 5009185), the estimated global market for industrial enzymes were USD 10.0 billion in 2019, which is continuously increasing as it is expected to reach about USD 14.7 billion by 2022. Among all enzymes, cellulases are the major group of enzymes act synergistically in breakdown of cellulose, that facilitates its conversion to various value-added products and also offer several other important applications at industrial scale. The hyper production of cellulases are required to overcome their demand of global market. Cellulases production can be enhanced by strain improvement as well as using advance fermentation technology. In this review a detail studies of strategies to enhance production of cellulases and improve their physiochemical properties for industrial application have been described.