Climate savvy grazing (Developing improved grazing and related practices to assist beef production enterprises across northern Australia to adapt to a changing and more variable climate)

There is uncertainty over the potential changes to rainfall across northern Australia under climate change. Since rainfall is a key driver of pasture growth, cattle numbers and the resulting animal productivity and beef business profitability, the ability to anticipate possible management strategies within such uncertainty is crucial. The Climate Savvy Grazing project used existing research, expert knowledge and computer modelling to explore the best-bet management strategies within best, median and worse-case future climate scenarios. All three scenarios indicated changes to the environment and resources upon which the grazing industry of northern Australia depends. Well-adapted management strategies under a changing climate are very similar to best practice within current climatic conditions. Maintaining good land condition builds resource resilience, maximises opportunities under higher rainfall years and reduces the risk of degradation during drought and failed wet seasons. Matching stocking rate to the safe long-term carrying capacity of the land is essential; reducing stock numbers in response to poor seasons and conservatively increasing stock numbers in response to better seasons generally improves profitability and maintains land in good condition. Spelling over the summer growing season will improve land condition under a changing climate as it does under current conditions. Six regions were included within the project. Of these, the Victoria River District in the Northern Territory, Gulf country of Queensland and the Kimberley region of Western Australia had projections of similar or higher than current rainfall and the potential for carrying capacity to increase. The Alice Springs, Maranoa-Balonne and Fitzroy regions had projections of generally drying conditions and the greatest risk of reduced pasture growth and carrying capacity. Encouraging producers to consider and act on the risks, opportunities and management options inherent in climate change was a key goal of the project. More than 60,000 beef producers, advisors and stakeholders are now more aware of the management strategies which build resource resilience, and that resilience helps buffer against the effects of variable and changing climatic conditions. Over 700 producers have stated they have improved confidence, skills and knowledge to attempt new practices to build resilience. During the course of the project, more than 165 beef producers reported they have implemented changes to build resource and business resilience.

Production of 2-phenethyl alcohol and 2-phenyllactic acid in Candida species

2-Phenethyl alcohol (2-PEA) and 2-phenyllactic acid (2-PLA) were isolated from the culture filtrates of Candida species grown in media containing peptone or phenylalanine as nitrogen source. These compounds were characterized by comparing their UV, IR, and NMR spectral properties with authentic samples. Candida species differed markedly in their production of 2-PEA and 2-PLA. Experiments using [14C]-phenylalanine indicated that both 2-PEA and 2-PLA are synthesised from L-phenylalanine. A pathway for the biosynthesis of 2-PEA from L-phenylalanine has been proposed.

Evaluation of tomato production systems at Bowen in mitigating the risk of fruit fly infestation: a systems approach for market access

Queensland fruit flies Bactrocera tryoni and B. neohumeralis are considered major quarantine pests of tomato, a major crop in the horticultural production district around Bowen, North Queensland, Australia. Preharvest and/or postharvest treatments are required to meet the market access requirements of both domestic and international trading partners. The suspension from use of dimethoate and fenthion, the two insecticides used for fruit fly control, has resulted in the loss of both pre and postharvest uses in fresh tomato. Research undertaken quantitatively at Bowen evaluated the effectiveness of pre-harvest production systems without specific fruit fly controls and postharvest mitigation measures in reducing the risk of fruit fly infestation in tomato. A district-wide trapping using cue-lure baited traps was undertaken to determine fruit fly seasonal patterns in relation to the cropping seasons. A total of 17,626 field-harvested and 11,755 pack-house tomatoes were sampled from ten farms over three cropping seasons (2006-2009). The fruit were incubated and examined for fruit fly infestation. No fruit fly infested fruit were recorded over the three seasons in either the field or the pack-house samples. Statistical analyses showed that upper infestation levels were extremely low (between 0.025 and 0.062%) at the 95% confidence level. The trap catches showed a seasonal pattern in fruit fly activity, with low numbers during the autumn and winter months, rising slightly in spring and peaking in summer. This seasonal pattern was similar over the four seasons. The main two species of fruit fly caught were B. tryoni and B. neohumeralis. Based on the results, it is clear that the risk of fruit fly infestation is extremely low under the current production systems in the Bowen region.

Developing Jungle Perch Fingerling Production to Improve Fishing Opportunities

This project has for the first time demonstrated the feasibility of hatchery production of jungle perch fingerlings. The research on jungle perch production has enabled a hatchery production manual with accompanying videos to be produced. This has given private commercial hatcheries the information needed to produce jungle perch fingerlings. Several hatcheries have already indicated an interest in producing jungle perch and will be assisted to do so in 2016. Currently jungle perch are not a permitted stocking species, so cannot be sold to fish stocking groups. However, hatcheries will be able to sell fingerlings to the aquarium trade or supply grow out facilities that could produce jungle perch for human consumption. Should jungle perch become a permitted species for stocking, this will provide hatcheries with a major new product option to sell to fish stocking groups. It would also benefit anglers by providing another iconic species for impoundment stocking programs. This could have flow-on benefits to regional economies through angler tourism. Should the pilot reintroductions of jungle perch into streams result in self-sustaining jungle perch populations, then there will be three restored jungle perch populations close to major population centres. This will create a new opportunity for anglers not normally able to target jungle perch. Since the majority of anglers who target jungle perch are catch and release fishers, angling is expected to have minimal impact on recovery of the populations. This project led to the development of a hatchery manual for jungle perch production and to a summary brochure. In late 2014 and in 2015 researchers were able to make the first ever releases of jungle perch fingerlings back into rivers and streams within their historical range.

The multidimensional causal factors of ‘wet litter’ in chicken-meat production

The problem of ‘wet litter’, which occurs primarily in grow-out sheds for meat chickens (broilers), has been recognised for nearly a century. Nevertheless, it is an increasingly important problem in contemporary chicken-meat production as wet litter and associated conditions, especially footpad dermatitis, have developed into tangible welfare issues. This is only compounded by the market demand for chicken paws and compromised bird performance. This review considers the multidimensional causal factors of wet litter. While many causal factors can be listed it is evident that the critical ones could be described as micro-environmental factors and chief amongst them is proper management of drinking systems and adequate shed ventilation. Thus, this review focuses on these environmental factors and pays less attention to issues stemming from health and nutrition. Clearly, there are times when related avian health issues of coccidiosis and necrotic enteritis cannot be overlooked and the development of efficacious vaccines for the latter disease would be advantageous. Presently, the inclusion of phytate-degrading enzymes in meat chicken diets is routine and, therefore, the implication that exogenous phytases may contribute to wet litter is given consideration. Opinion is somewhat divided as how best to counter the problem of wet litter as some see education and extension as being more beneficial than furthering research efforts. However, it may prove instructive to assess the practice of whole grain feeding in relation to litter quality and the incidence of footpad dermatitis. Additional research could investigate the relationships between dietary concentrations of key minerals and the application of exogenous enzymes with litter quality.

Comparison of the impact of six heat-load management strategies on thermal responses and milk production of feed-pad and pasture fed dairy cows in a subtropical environment

Exposure to hot environments affects milk yield (MY) and milk composition of pasture and feed-pad fed dairy cows in subtropical regions. This study was undertaken during summer to compare MY and physiology of cows exposed to six heat-load management treatments. Seventy-eight Holstein-Friesian cows were blocked by season of calving, parity, milk yield, BW, and milk protein (%) and milk fat (%) measured in 2 weeks prior to the start of the study. Within blocks, cows were randomly allocated to one of the following treatments: open-sided iron roofed day pen adjacent to dairy (CID) + sprinklers (SP); CID only; non-shaded pen adjacent to dairy + SP (NSD + SP); open-sided shade cloth roofed day pen adjacent to dairy (SCD); NSD + sprinkler (sprinkler on for 45 min at 1100 h if mean respiration rate >80 breaths per minute (NSD + WSP)); open-sided shade cloth roofed structure over feed bunk in paddock + 1 km walk to and from the dairy (SCP + WLK). Sprinklers for CID + SP and NSD + SP cycled 2 min on, 12 min off when ambient temperature >26°C. The highest milk yields were in the CID + SP and CID treatments (23.9 L cow−1 day−1), intermediate for NSD + SP, SCD and SCP + WLK (22.4 L cow−1 day−1), and lowest for NSD + WSP (21.3 L cow−1 day−1) (P < 0.05). The highest (P < 0.05) feed intakes occurred in the CID + SP and CID treatments while intake was lowest (P < 0.05) for NSD + WSP and SCP + WLK. Weather data were collected on site at 10-min intervals, and from these, THI was calculated. Nonlinear regression modelling of MY × THI and heat-load management treatment demonstrated that cows in CID + SP showed no decline in MY out to a THI break point value of 83.2, whereas the pooled MY of the other treatments declined when THI >80.7. A combination of iron roof shade plus water sprinkling throughout the day provided the most effective control of heat load.

Jungle Perch Kuhlia Rupestris Fingerling Production Manual

This manual consists of written descriptions of jungle perch Kuhlia rupestris production and video material to demonstrate each of the key production steps. Video links are at the end of each major written section in the document. To activate the link use ctrl click. The videos enhance the instructive ability of this manual. The keys to producing jungle perch are:  maintaining broodstock in freshwater or low salinity water less than 5 ppt  spawning fish in full seawater at 28C  incubating eggs in full seawater. Salinities must not be less than 32 ppt  ensuring that first feed jungle perch larvae have an adequate supply of copepod nauplii  rearing larvae in full seawater under bright light  use of gentle aeration in tanks  postponing spawns until adequate densities of copepod nauplii are present in ponds  sustaining copepod blooms in ponds for at least 20 days  avoiding use of paddlewheels in ponds  supplementary feeding with Artemia salina and weaning diets from 20 days after hatch  harvesting of fingerlings or fry after they are 25-30 mm in length (50 to 60 days post hatch)  covering tanks of fingerlings with 5 mm mesh and submerging freshwater inlets to prevent jumping.

Production of cast aluminium-graphite particle composites using a pellet method

Copper- and nickel-coated graphite particles can be successfully introduced into aluminium-base alloy melts as pellets to produce cast aluminium-graphite particle composites. The pellets were made by pressing mixtures of nickel- or copper-coated graphite particles and aluminium powders together at pressures varying between 2 and 20 kg mm–2. These pellets were dispersed in aluminium alloy melts by plunging and holding them in the melts using a refractory coated mild steel cone, until the pellets disintegrated and the powders were dispersed. The optimum pressure for the preparation of pellets was 2 to 5 kg mm–2 and the optimum size and percentage of aluminium powder were 400 to 1000mgrm and 35 wt% respectively. Under optimum conditions the recovery of the graphite particles in the castings was as high as 96%, these particles being pushed into the last freezing interdendritic regions. The tensile strength and the hardness of the graphite aluminium alloys made using the pellet method are comparable to those of similar composites made using gas injection or the vortex method. The pellet method however has the advantage of greater reproducibility and flexibility. Dispersion of graphite particles in the matrix of cast aluminium alloys using the pellet method increases their resistance to wear.

Dental Service Utilization, Dental Health Production and Equity in Dental Care: the Finnish Experience

This thesis is grounded on four articles. Article I generally examines the factors affecting dental service utilization. Article II studies the factors associated with sector-specific utilization among young adults entitled to age-based subsidized dental care. Article III explores the determinants of dental ill-health as measured by the occurrence of caries and the relationship between dental ill-health and dental care use. Article IV measures and explains income-related inequality in utilization. Data employed were from the 1996 Finnish Health Care Survey (I, II, IV) and the 1997 follow-up study included in the longitudinal study of the Northern Finland 1966 Birth Cohort (III). Utilization is considered as a multi-stage decision-making process and measured as the number of visits to the dentist. Modified count data models and concentration and horizontal equity indices were applied. Dentist s recall appeared very efficient at stimulating individuals to seek care. Dental pain, recall, and the low number of missing teeth positively affected utilization. Public subvention for dental care did not seem to statistically increase utilization. Among young adults, a perception of insufficient public service availability and recall were positively associated with the choice of a private dentist, whereas income and dentist density were positively associated with the number of visits to private dentists. Among cohort females, factors increasing caries were body mass index and intake of alcohol, sugar, and soft drinks and those reducing caries were birth weight and adolescent school achievement. Among cohort males, caries was positively related to the metropolitan residence and negatively related to healthy diet and education. Smoking increased caries, whereas regular teeth brushing, regular dental attendance and dental care use decreased caries. We found equity in young adults utilization but pro-rich inequity in the total number of visits to all dentists and in the probability of visiting a dentist for the whole sample. We observed inequity in the total number of visits to the dentist and in the probability of visiting a dentist, being pro-poor for public care but pro-rich for private care. The findings suggest that to enhance equal access to and use of dental care across population and income groups, attention should focus on supply factors and incentives to encourage people to contact dentists more often. Lowering co-payments and service fees and improving public availability would likely increase service use in both sectors. To attain favorable oral health, appropriate policies aimed at improving dental health education and reducing the detrimental effects of common risk factors on dental health should be strengthened. Providing equal access with respect to need for all people ought to take account of the segmentation of the service system, with its two parallel delivery systems and different supplier incentives to patients and dentists.

An operational-level multi-stage mine production timetabling model for optimally synchronising drilling, blasting and excavating operations

This paper proposes a new multi-stage mine production timetabling (MMPT) model to optimise open-pit mine production operations including drilling, blasting and excavating under real-time mining constraints. The MMPT problem is formulated as a mixed integer programming model and can be optimally solved for small-size MMPT instances by IBM ILOG-CPLEX. Due to NP-hardness, an improved shifting-bottleneck-procedure algorithm based on the extended disjunctive graph is developed to solve large-size MMPT instances in an effective and efficient way. Extensive computational experiments are presented to validate the proposed algorithm that is able to efficiently obtain the near-optimal operational timetable of mining equipment units. The advantages are indicated by sensitivity analysis under various real-life scenarios. The proposed MMPT methodology is promising to be implemented as a tool for mining industry because it is straightforwardly modelled as a standard scheduling model, efficiently solved by the heuristic algorithm, and flexibly expanded by adopting additional industrial constraints.

Phylogeography and Population Genetics of the Moor frog (Rana arvalis Nilsson) in Northern Europe

The ongoing climate change along with increasing levels of pollutants, diseases, habitat loss and fragmentation constitute global threats to the persistence of many populations, species and ecosystems. However, for the long-term persistence of local populations, one of the biggest threats is the intrinsic loss of genetic variation. In order to adapt to changes in the environment, organisms must have a sufficient supply of heritable variation in traits important for their fitness. With a loss of genetic variation, the risk of extinction will increase. For conservational practices, one should therefore understand the processes that shape the genetic population structure and also the broader (historical) phylogenetic patterning of the species in focus. In this thesis, microsatellite markers were applied to study genetic diversity and population differentiation of the protected moor frog (Rana arvalis) in Fennoscandia from both historical (evolutionary) and applied (conservation) perspectives. The results demonstrate that R. arvalis populations are highly structured over rather short geographic distances. Moreover, the results suggest that R. arvalis recolonized Fennoscandia from two directions after the last ice age. This has had implications for the genetic structuring and population differentiation, especially in the northernmost parts where the two lineages have met. Compared to more southern populations, the genetic variation decreases and the interpopulation differentiation increases dramatically towards north. This could be an outcome of serial population bottlenecking along the recolonization route. Also, current isolation and small population sizes increase the effect of drift, thus reinforcing the observed pattern. The same pattern can also be seen in island populations. However, though R. arvalis on the island of Gotland has lost most of its neutral genetic variability, our results indicate that the levels of additive genetic variation have remained high. This conforms to the conjecture that though neutral markers are widely used in conservation purposes, they may be quite uninformative about the levels of genetic variation in ecologically important traits. Finally, the evolutionary impact of the typical amphibian mating behaviour on genetic diversity was investigated. Given the short time available for larval development, it is important that mating takes place as early as possible. The genetic data and earlier capture-recapture data suggest that R. arvalis gather at mating grounds they are familiar with. However, by forming leks in random to relatedness, and having multiple paternities in single clutches, the risk of inbreeding may be minimized in this otherwise highly philopatric species.

GDNF family receptors in peripheral target innervation and hormone production

Glial cell line-derived neurotrophic factor (GDNF) family ligands: GDNF, neurturin, persephin and artemin, signal through a receptor tyrosine kinase Ret by binding first to a co-receptor (GFRα1-4) that is attached to the plasma membrane. The GDNF family factors can support the survival of various peripheral and central neuronal populations and have important functions also outside the nervous system, especially in kidney development. Activating mutations in the RET gene cause tumours in neuroendocrine cells, whereas inactivating mutations in RET are found in patients with Hirschsprung s disease (HSCR) characterized by loss of ganglionic cells along the intestine. The aim of this study was to examine the in vivo functions of neurturin receptor GFRα2 and persephin receptor GFRα4 using knockout (KO) mice. Mice lacking GFRα2 grow poorly after weaning and have deficits in parasympathetic and enteric innervation. This study shows that impaired secretion of the salivary glands and exocrine pancreas contribute to growth retardation in GFRα2-KO mice. These mice have a reduced number of intrapancreatic neurons and decreased cholinergic innervation of the exocrine pancreas as well as reduced excitatory fibres in the myenteric plexus of the small intestine. This study also demonstrates that GFRα2-mediated Ret signalling is required for target innervation and maintenance of soma size of sympathetic cholinergic neurons and sensory nociceptive IB4-binding neurons. Furthermore, lack of GFRα2 in mice results in deficient perception of temperatures above and below thermoneutrality and in attenuated inflammatory pain response. GFRα4 is co-expressed with Ret predominantly in calcitonin-producing thyroid C-cells in the mouse. In this study GFRα4-deficient mice were generated. The mice show no gross developmental deficits and have a normal number of C-cells. However, young but not adult mice lacking GFRα4 have a lower production of calcitonin in thyroid tissue and consequently, an increased bone formation rate. Thus, GFRα4/Ret signalling may regulate calcitonin production. In conclusion, this study reveals that GFRα2/Ret signalling is crucial for the development and function of specific components of the peripheral nervous system and that GFRα4-mediated Ret signalling is required for controlling transmitter synthesis in thyroid C-cells.

Pentitol phosphate dehydrogenases: Discovery, characterization and use in D-arabitol and xylitol production by metabolically engineered Bacillus subtilis

The ultimate goal of this study has been to construct metabolically engineered microbial strains capable of fermenting glucose into pentitols D-arabitol and, especially, xylitol. The path that was chosen to achieve this goal required discovery, isolation and sequencing of at least two pentitol phosphate dehydrogenases of different specificity, followed by cloning and expression of their genes and characterization of recombinant arabitol and xylitol phosphate dehydrogenases. An enzyme of a previously unknown specificity, D-arabitol phosphate dehydrogenase (APDH), was discovered in Enterococcus avium. The enzyme was purified to homogenity from E. avium strain ATCC 33665. SDS/PAGE revealed that the enzyme has a molecular mass of 41 ± 2 kDa, whereas a molecular mass of 160 ± 5 kDa was observed under non-denaturing conditions implying that the APDH may exist as a tetramer with identical subunits. Purified APDH was found to have narrow substrate specificity, converting only D-arabitol 1-phosphate and D-arabitol 5-phosphate into D-xylulose 5-phosphate and D-ribulose 5-phosphate, respectively, in the oxidative reaction. Both NAD+ and NADP+ were accepted as co-factors. Based on the partial protein sequences, the gene encoding APDH was cloned. Homology comparisons place APDH within the medium chain dehydrogenase family. Unlike most members of this family, APDH requires Mn2+ but no Zn2+ for enzymatic activity. The DNA sequence surrounding the gene suggests that it belongs to an operon that also contains several components of phosphotransferase system (PTS). The apparent role of the enzyme is to participate in arabitol catabolism via the arabitol phosphate route similar to the ribitol and xylitol catabolic routes described previously. Xylitol phosphate dehydrogenase (XPDH) was isolated from Lactobacillus rhamnosus strain ATCC 15820. The enzyme was partially sequenced. Amino acid sequences were used to isolate the gene encoding the enzyme. The homology comparisons of the deduced amino acid sequence of L. rhamnosus XPDH revealed several similar enzymes in genomes of various species of Gram-positive bacteria. Two enzymes of Clostridium difficile and an enzyme of Bacillus halodurans were cloned and their substrate specificities together with the substrate specificity of L. rhamnosus XPDH were compared. It was found that one of the XPDH enzymes of C. difficile and the XPDH of L. rhamnosus had the highest selectivity towards D-xylulose 5-phosphate. A known transketolase-deficient and D-ribose-producing mutant of Bacillus subtilis (ATCC 31094) was further modified by disrupting its rpi (D-ribose phosphate isomerase) gene to create D-ribulose- and D-xylulose-producing strain. Expression of APDH of E. avium and XPDH of L. rhamnosus and C. difficile in D-ribulose- and D-xylulose-producing strain of B. subtilis resulted in strains capable of converting D-glucose into D-arabitol and xylitol, respectively. The D-arabitol yield on D-glucose was 38 % (w/w). Xylitol production was accompanied by co-production of ribitol limiting xylitol yield to 23 %.

Characterisation, cloning and production of industrially interesting enzymes: gluconolactone oxidase of Penicillium cyaneo-fulvum and gluconate 5-dehydrogenase of Gluconobacter suboxydans

The work covered in this thesis is focused on the development of technology for bioconversion of glucose into D-erythorbic acid (D-EA) and 5-ketogluconic acid (5-KGA). The task was to show on proof-of-concept level the functionality of the enzymatic conversion or one-step bioconversion of glucose to these acids. The feasibility of both studies to be further developed for production processes was also evaluated. The glucose - D-EA bioconversion study was based on the use of a cloned gene encoding a D-EA forming soluble flavoprotein, D-gluconolactone oxidase (GLO). GLO was purified from Penicillium cyaneo-fulvum and partially sequenced. The peptide sequences obtained were used to isolate a cDNA clone encoding the enzyme. The cloned gene (GenBank accession no. AY576053) is homologous to the other known eukaryotic lactone oxidases and also to some putative prokaryotic lactone oxidases. Analysis of the deduced protein sequence of GLO indicated the presence of a typical secretion signal sequence at the N-terminus of the enzyme. No other targeting/anchoring signals were found, suggesting that GLO is the first known lactone oxidase that is secreted rather than targeted to the membranes of the endoplasmic reticulum or mitochondria. Experimental evidence supports this analysis, as near complete secretion of GLO was observed in two different yeast expression systems. Highest expression levels of GLO were obtained using Pichia pastoris as an expression host. Recombinant GLO was characterised and the suitability of purified GLO for the production of D-EA was studied. Immobilised GLO was found to be rapidly inactivated during D-EA production. The feasibility of in vivo glucose - D-EA conversion using a P. pastoris strain co-expressing the genes of GLO and glucose oxidase (GOD, E.C. 1.1.3.4) of A. niger was demonstrated. The glucose - 5-KGA bioconversion study followed a similar strategy to that used in the D-EA production research. The rationale was based on the use of a cloned gene encoding a membrane-bound pyrroloquinoline quinone (PQQ)-dependent gluconate 5-dehydrogenase (GA 5-DH). GA 5-DH was purified to homogeneity from the only source of this enzyme known in literature, Gluconobacter suboxydans, and partially sequenced. Using the amino acid sequence information, the GA 5-DH gene was cloned from a genomic library of G. suboxydans. The cloned gene was sequenced (GenBank accession no. AJ577472) and found to be an operon of two adjacent genes encoding two subunits of GA 5-DH. It turned out that GA 5-DH is a rather close homologue of a sorbitol dehydrogenase from another G. suboxydans strain. It was also found that GA 5-DH has significant polyol dehydrogenase activity. The G. suboxydans GA 5-DH gene was poorly expressed in E. coli. Under optimised conditions maximum expression levels of GA 5-DH did not exceed the levels found in wild-type G. suboxydans. Attempts to increase expression levels resulted in repression of growth and extensive cell lysis. However, the expression levels were sufficient to demonstrate the possibility of bioconversion of glucose and gluconate into 5-KGA using recombinant strains of E. coli. An uncharacterised homologue of GA 5-DH was identified in Xanthomonas campestris using in silico screening. This enzyme encoded by chromosomal locus NP_636946 was found by a sequencing project of X. campestris and named as a hypothetical glucose dehydrogenase. The gene encoding this uncharacterised enzyme was cloned, expressed in E. coli and found to encode a gluconate/polyol dehydrogenase without glucose dehydrogenase activity. Moreover, the X. campestris GA 5-DH gene was expressed in E. coli at nearly 30 times higher levels than the G. suboxydans GA 5-DH gene. Good expressability of the X. campestris GA-5DH gene makes it a valuable tool not only for 5-KGA production in the tartaric acid (TA) bioprocess, but possibly also for other bioprocesses (e.g. oxidation of sorbitol into L-sorbose). In addition to glucose - 5-KGA bioconversion, a preliminary study of the feasibility of enzymatic conversion of 5-KGA into TA was carried out. Here, the efficacy of the first step of a prospective two-step conversion route including a transketolase and a dehydrogenase was confirmed. It was found that transketolase convert 5-KGA into TA semialdehyde. A candidate for the second step was suggested to be succinic dehydrogenase, but this was not tested. The analysis of the two subprojects indicated that bioconversion of glucose to TA using X. campestris GA 5-DH should be prioritised first and the process development efforts in future should be focused on development of more efficient GA 5-DH production strains by screening a more suitable production host and by protein engineering.

Effects of glycerol on enzymatic hydrolysis and ethanol production using sugarcane bagasse pretreated by acidified glycerol solution

In this study, for the first time the effects of glycerol on enzymatic hydrolysis and ethanol fermentation were investigated. Enzymatic hydrolysis was inhibited slightly with 2.0 wt% glycerol, leading to reduction in glucan digestibility from 84.9% without glycerol to 82.9% (72 h). With 5.0 wt% and 10.0 wt% glycerol, glucan digestibility reduced by 4.5% and11.0%, respectively. However, glycerol appeared not detrimental to cellulase enzymes. Ethanol fermentation was not affected with glycerol up to 5.0 wt%, and was inhibited slightly with 10.0 wt% glycerol, which resulted in reduction in ethanol yield from 86.0% without glycerol to 83.7% (20 h). Based on laboratory and pilot scale enzymatic hydrolysis and ethanol production results, it was estimated that 0.142 kg ethanol could be produced from 1.0 kg dry bagasse (a glucan content of 38.0%) after pretreatment with acidified glycerol solution.