Debranching enzyme concentration effected on physicochemical properties and α-amylase hydrolysis rate of resistant starch type III from amylose rice starch

The effect of debranching enzyme concentration on physicochemical properties and α-amylase hydrolysis rate of resistant starch type III from high amylose rice starch were studied. The pullulanase enzyme (8, 10, 12, 14 and 16 U/g starch) was introduced to modify amylopectin molecules of 15% (w/w) gelatinized rice starches at 55 °C for 16 h. The debranched starches with different degrees of hydrolysis (0.14–5.27%), and having 66.60–98.82% β-amylolysis limit were then induced at 4 °C for 16 h, afterward a one cycle of freeze–thaw process (?10/30 °C) was applied. The results showed that a pullulanase hydrolysis improved the degree of syneresis (51.64–54.85% from 8 to 16 U/g starch). Resistant starch content increased sharply as the amount of the enzyme increased, reaching the highest (19.81%) for a 12 U/g starch and decreased to 13.16% by 16 U/g starch. α-Amylase hydrolysis rate showed that incompletely-debranched had a lower estimated glycemic index than completely debranched rice starches. Microstructure of the selected RS III samples using X-ray diffraction and scanning electron microscopy revealed a crystal pattern change from A- to V-type pattern and formed a coarse honeycomb-like and a filamentous network structure.     

Pasting properties of a heat-moisture treated canna starch in relation to its structural characteristics

Native and moistened canna starches (moisture contents of 15%, 18%, 20%, 22%, and 25%) were heat treated at 100 °C for 16 h. Heat-moisture treatment (HMT) did not alter the shape or size of starch granules. Heat-treated starches showed lower paste viscosity, more stable paste (with no breakdown) and noticeable decrease in setback value compared to untreated native starch. These changes were more obvious when the starch samples contained more moisture. Starch gel morphology investigated by I₂ staining revealed that the granules of untreated native canna starch were completely ruptured, whereas those of HMT22% and HMT25% samples remained in granular form. These gel morphologies were correlated and provided the information to explain the pasting behaviors of treated and untreated canna starches. Shifts of gelatinization endotherms towards higher temperatures (1–8 °C) with broadened peaks following the moisture contents in starch samples were found after HMT. Amylose leaching of treated samples tended to be lower when the moisture content of the samples increased. HMT did not alter crystalline type of canna starch (B-type), but with increase of the moisture, slight reduction of a peak at 5.7° and a fusion of doublet at 22° and 24° were observed.     

Impact of annealing and heat-moisture treatment on rapidly digestible,slowly digestible and resistant starch levels in native and gelatinized corn,pea and lentil starches

Impact of annealing (ANN) and heat-moisture treatment (HMT) on rapidly digestible starch (RDS), slowly digestible starch (SDS), resistant starch (RS), and expected glycemic index (eGI) of corn, pea, and lentil starches in their native and gelatinized states were determined. ANN was done for 24 h at 70% moisture at temperatures 10 and 15 °C below the onset (T_o) temperature of gelatinization, while HMT was done at 30% moisture at 100 and 120 °C for 2 h. The swelling factor (SF), amylose leaching (AML) and gelatinization parameters of the above starches before and after ANN and HMT were determined. SF and AML decreased on ANN and HMT (HMT > ANN). The gelatinization temperatures increased on ANN and HMT (HMT > ANN). However, the gelatinization temperature range decreased on ANN but increased on HMT. Birefringence remained unchanged on ANN but decreased on HMT. The Fourier transform infrared (FT-IR) absorbance ratio of 1047 cm^?1/1022 cm^?1 increased on ANN but decreased on HMT. ANN and HMT increased RDS, RS and eGI levels and decreased SDS levels in granular starches. HMT had a greater impact than ANN on RDS, RS, and SDS levels. In gelatinized starches, ANN and HMT decreased RDS and eGI, but increased SDS and RS levels. These changes were more pronounced on HMT. This study showed that amylopectin structure and interactions formed during ANN and HMT had a significant impact on RDS, SDS, RS and eGI levels of starches.     

A comparative study of some properties of cassava (Manihot esculenta,Crantz) and cocoyam (Colocasia esculenta,Linn) starches

Some properties of cassava and cocoyam starches were studied and compared with a view to determining the functional applications in food systems for which they are suitable. The starches were compared in terms of their microscopic, thermal, physicochemical and rheological properties. Microscopy revealed smaller granule sizes of cocoyam starch compared with cassava. The amylose content was higher in cocoyam starch (33.3%) than in cassava starch (29.3%). Gelatinization in cassava starch occurred at a lower temperature range (60.11–72.67 °C) compared with cocoyam (72.96–80.25 °C) with the endothermic gelatinization enthalpy higher in cocoyam. The swelling power and solubility patterns indicated lower relaxation temperature, higher swelling and solubilization rates in cassava starch compared with cocoyam starch. The pasting characteristics of 8% (db, dry basis) starch slurry showed that cassava had higher peak viscosity but lower stability and setback ratios compared with cocoyam. This indicates that cocoyam starch paste is better in withstanding processing conditions and would present a superior thickening characteristic than cassava starch paste. The flow properties of both starch pastes showed non-Newtonian behaviour and could be best described by the Herschel–Bulkley model. The rate index and yield stress of cocoyam starch paste was higher than that of cassava. The storage modulus of cocoyam starch paste was higher than that of cassava indicating that cocoyam starch paste was more rigid than cassava starch paste. Cassava starch paste exhibited higher paste clarity and freeze-thaw stability than cocoyam starch paste. The properties of cassava and cocoyam starches dictate their food applications.     

Origin of defects in assembled supramolecular structures of sweet potato starches with different amylopectin chain-length distribution

A combined approach of fluorophore-assisted capillary electrophoresis (FACEL), high-sensitivity differential scanning calorimetry (DSC), wide-angle X-ray scattering (WAXS), small-angle X-ray scattering (SAXS), and light (LM) and scanning electron microscopy (SEM) was applied to study the effects of changes in amylopectin chain-length distribution on the assembly structures of sweet potato starches with similar amylose levels. It was shown that unlike ordinary sweet potato starch, starch extracted from Quick Sweet cultivar of sweet potato had anomalous high level of amylopectin chains with a degree of polymerization (DP) 6–12. Joint analysis of the obtained data revealed that amylopectin chains with DP 10–24 are, apparently, the dominant material for the formation of supramolecular structures in starch granules. In contrast, amylopectin chains with DP < 10 facilitated the formation of defects within crystalline lamellae. An increase in relative content of amylopectin chains with DP < 10 is accompanied by the correlated structural alterations manifested at all levels of starch granule organization (crystalline lamellae, amylopectin clusters, semi-crystalline growth rings, and granule morphology). Thus, the short amylopectin chains with DP < 10 were considered as an origin of the defectiveness in starch supramolecular structures.     

Some properties of white and yellow plantain (Musa paradisiaca,Normalis) starches

Starch obtained from yellow and white plantain varieties were subjected to proximate analysis, physicochemical and rheological characterization in order to evaluate their properties. Yellow plantain variety gave higher yield of starch than the white variety. The two varieties differed in the purity of starch extract; white plantain starch contained: ash (1.09%), protein (0.640%) and fat (0.276%) while yellow plantain starch contained: ash (0.95%), protein (0.325%) and fat (0.403%). The amylose content of yellow plantain starch (24.36% (apparent), 26.13% (total)) was similar to that of white plantain starch (24.24% (apparent), 26.01% (total)). Scanning electron microscopy revealed bimodal irregular shaped granules (3.74–7.00 and 10.00–33.00 μm) in white plantain starch and elliptical granules (11.22–41.00 μm) in yellow plantain starch. Both starches differed markedly in their physicochemical properties. Their differences in gelatinization temperature (yellow plantain, 64.99–73.90 °C; white plantain, 68.08–77.15 °C), swelling and solubility patterns, and pasting characteristics indicated that yellow plantain starch had weaker granule architecture compared with white plantain starch. Further evidence of differences in properties was obtained from flow and viscoelastic properties of the starch gels, paste clarity and freeze–thaw stability.     

Optimization of the synthesis of 1-allyloxy-2-hydroxy-propyl-starch through statistical experimental design

The synthesis of 1-allyloxy-2-hydroxy-propyl starches was studied using a statistical experimental design approach. The etherification of two different granular maize starches with allyl glycidyl ether (AGE) in a heterogeneous alkaline suspension was investigated. The optimal reaction conditions were found via experimental design and the obtained response factor, e.g. the degree of substitution (DS) of the starch hydroxyl group, was statistically evaluated. The effects of six process factors on DS, namely the starch concentration, the reaction time, the temperature, and the amount of NaOH, Na₂SO₄ and AGE were investigated. The statistical analysis showed significant impact of the temperature, the amount of NaOH and the amount of AGE on the DS for both starches. Optimum conditions for the highest DS for waxy maize starch were: 0.0166% AGE (based on dry starch (ds)) and 1.0% NaOH (ds) at 34 °C in 4 h; on dent maize starch, these were 0.0099% AGE (ds) and 1.0% NaOH (ds) at 37 °C in 16 h.     

The impact of single and dual hydrothermal modifications on the molecular structure and physicochemical properties of normal corn starch

Effect of single and dual hydrothermal modifications with annealing (ANN) and heat-moisture treatment (HMT) on molecular structure and physicochemical properties of corn starch was investigated. Normal corn starch was modified by ANN at 70% moisture at 50 °C for 24 h and HMT at 30% moisture at 120 °C for 24 h as well as by the combination of ANN and HMT. The apparent amylose content and swelling factor (SF) decreased on ANN and HMT, but amylose leaching (AML) increased. These changes were more pronounced on dual modification. The crystallinity (determined by X-ray diffraction), the gelatinization enthalpy (determined by differential scanning calorimetry) and ratio of 1047 cm^?1/1022 cm^?1 (determined by Fourier transform infrared spectroscopy) slightly increased on ANN and decreased on HMT. The ANN and subsequent HMT (ANN-HMT) resulted in the lowest crystallinity, gelatinization enthalpy and ratio of 1047 cm^?1/1022 cm^?1. The gelatinization temperature range decreased on ANN but increased on HMT. However, the gelatinization range of dually modified starches (ANN-HMT and HMT-ANN) was between ANN starch and HMT starch. Birefringence remained unchanged on ANN but slightly decreased on HMT as well as dual modification. Average chain length and amount of longer branch chains (DP ≥ 37) remained almost unchanged on ANN but decreased on HMT and dual modifications (ANN-HMT and HMT-ANN). HMT and dual modifications resulted in highly reduced pasting viscosity. ANN and HMT as well as dual modifications increased RDS content and decreased SDS and RS content.     

Starch pastes thinning during high-pressure homogenization

High-pressure homogenization induced thinning of potato and cassava starch paste was investigated. The starch slurries at a concentration of 2.0 wt.% were heated at 90 °C for 1 h and then rapidly cooled in tap water. The cooled starch pastes were homogenized at various pressures ranging from 0 to 100 MPa using a lab-scale high-pressure homogenizer. The influence of homogenizing pressure on the temperature, apparent viscosity, electrical conductivity, and percent light transmittance of homogenized starch pastes were determined. Temperatures of homogenized starch pastes increased linearly with the increase of the applied pressure, and the rate was 0.177 °C/MPa and 0.186 °C/MPa for potato and cassava starch pastes, respectively. After high-pressure homogenization, the apparent viscosities of the starch pastes decreased, while the percent light transmittances of them increased. However, the electrical conductivities of starch pastes were not affected by homogenization.     

Effect of the cross-linked reagent type on some morphological,physicochemical and functional characteristics of banana starch (Musa paradisiaca)

Cross-linked starches have increased their importance due to their applications such as adsorbents of heavy metals. In this work the effect the reagent used in the chemical modification of banana starch and its impact on some morphological, physicochemical and functional characteristics was evaluated. The reagent used in the cross-linked of starch decreased the fat and protein content, whereas ash level were higher. The morphology of the granules, observed by scanning electron microscopy, was more affected when a blend of sodium trimetaphosphate (STMP)/sodium tripolyphosphate (STPP) and epichlorohydrin (EPI) were used in the modification. The cross-linked starches presented a bimodal distribution and the effect was more conspicuous in those starches modified with STMP/STPP and EPI. The swelling value (60 °C) increased with the cross-linking and the highest value was obtained in those starches modified with STMP/STPP and EPI. However, at higher temperatures the swelling values of cross-linked starches with STMP/STPP and EPI decreased as temperature increased (80 °C), and there after the value was constant. The cross-linked starches with STMP/STPP and EPI showed the lowest solubility values. The cross-linked starch with POCl₃ (phosphorous oxychloride) showed a slight decrease in the onset and peak temperatures compared with its native counterpart, but those modified with STMP/STPP and EPI presented an increase in the three transition temperatures, but a decrease in enthalpy value. The results obtained can be used to determine the type of reagent used for cross-linked in order to obtain a starch with specific characteristics.     

Purification and biochemical characterization of a novel SDS and surfactant stable,raw starch digesting,and halophilic α-amylase from a moderately halophilic bacterium,Nesterenkonia sp. strain F

An extracellular halophilic α-amylase from Nesterenkonia sp. strain F was purified to homogeneity by 80% ethanol precipitation, Q-Sepharose anion exchange and Sephacryl S-200 gel filtration chromatography, with a 10.8-fold increase in specific activity. The molecular mass of the amylase was estimated to be 100 kDa and 106 kDa by SDS–PAGE and gel filtration chromatography, respectively. The enzyme showed maximal activity at pH 7.5 and 45 °C. The amylase was active in a wide range of salt concentrations (0–4 M) with its maximum activity at 0.5 M NaCl or 1 M KCl and was stable at the salts concentrations between 1 M and 4 M. Fe³⁺, Cu²⁺, Zn²⁺ and Al³⁺ strongly inhibited the enzyme, whereas Ca²⁺ stimulated the amylase activity. The α-amylase was inhibited by EDTA, but was not inhibited by PMSF and β-mercaptoethanol. The enzyme showed remarkable stability towards 0.5% SDS and sarcosyl, and 2% each of Triton X-100, Tween 80 and Tween 20. K_m value of the amylase for soluble starch was 4.5 mg/ml. The amylase hydrolyzed 38% of raw wheat starch and 20% of corn starch in a period of 48 h. The major products of soluble starch hydrolysis were maltose, maltotriose and maltotetraose, indicating an α-amylase activity.     

α-Amylase from germinating soybean (Glycine max) seeds – Purification,characterization and sequential similarity of conserved and catalytic amino acid residues

Starch hydrolyzing amylase from germinated soybeans seeds (Glycine max) has been purified 400-fold to electrophoretic homogeneity with a final specific activity of 384 units/mg. SDS–PAGE of the final preparation revealed a single protein band of 100 kDa, whereas molecular mass was determined to be 84 kDa by MALDI–TOF and gel filtration on Superdex-200 (FPLC). The enzyme exhibited maximum activity at pH 5.5 and a pI value of 4.85. The energy of activation was determined to be 6.09 kcal/mol in the temperature range 25–85 °C. Apparent Michaelis constant (K_m(app)) for starch was 0.71 mg/mL and turnover number (k_cat) was 280 s^?1 in 50 mM sodium acetate buffer, pH 5.5. Thermal inactivation studies at 85 °C showed first-order kinetics with rate constant (k) equal to 0.0063 min^?1. Soybean α-amylase showed high specificity for its primary substrate starch. High similarity of soybean α-amylase with known amylases suggests that this α-amylase belongs to glycosyl hydrolase family 13. Cereal α-amylases have gained importance due to their compatibility for biotechnological applications. Wide availability and easy purification protocol make soybean as an attractive alternative for plant α-amylase. Soybean can be used as commercially viable source of α-amylase for various industrial applications.     

Leaf non-structural carbohydrates regulated by plant functional groups and climate: Evidences from a tropical to cold-temperate forest transect

Non-structural carbohydrates (NSCs), e.g., glucose and starch, play important roles in metabolic processes of plants and represent important functional traits in plant's adaptation to external environment. To explore the variations in leaf NSCs among species and communities at a large scale and their influencing factors, we investigated the contents of leaf NSCs among 890 plant species in nine typical forests along the north–south transect of eastern China. The results showed that the contents of leaf soluble sugars, starch, and NSCs (sugars + starch) were highly variable among different plant species on the site scale, and their mean values for the 890 plant species were 45.7 mg g⁻¹, 47.5 mg g⁻¹, and 93.2 mg g⁻¹, respectively. All three metrics varied markedly across plant functional groups in the order of trees < shrubs < herbs. Weak latitudinal patterns of leaf soluble sugars, starch, and NSCs were observed from tropical to cold-temperate forests at the levels of species and plant functional groups. The contents of leaf soluble sugars, starch, and NSCs decreased with increasing temperature and precipitation which supports the growth limitation hypothesis at a large scale. In trees, leaf soluble sugars, starch, and NSCs increased with increasing photosynthetic active radiation (PAR); and were positively correlated with specific leaf area (SLA). The spatial patterns of leaf NSCs in forests along the north–south transect of eastern China and their relationships with temperature, precipitation, PAR, and SLA illustrate an important adaptation of plant communities to environmental changes at the continental scale.     

Phase state and surface topography of palmitoyl-ceramide monolayers

In cell biology (and in many biophysical) studies there is a natural tendency to consider ceramide as a highly condensed, solid-type lipid conferring rigidity and close packing to biomembranes. In the present work we advanced the understanding of the phase behavior of palmitoyl-ceramide restricted to a planar interface using Langmuir monolayers under strictly controlled and known surface packing conditions. Surface pressure–molecular area isotherms were complemented with molecular area–temperature isobars and with observations of the surface topography by Brewster Angle Microscopy. The results described herein indicate that palmitoyl-ceramide can exhibit expanded, as well as condensed phase states. Formation of three phases was found, depending on the surface pressure and temperature: a solid (1.80 nm thick), a liquid-condensed (1.73 nm thick, likely tilted) and a liquid-expanded (1.54 nm thick) phase over the temperature range 5–62 °C. A large hysteretic behavior is observed for the S phase monolayer that may indicate high resistance to domain boundary deformation. A second (or higher) order S → LC phase transition is observed at about room temperature while a first order LC → LE transition occurs in a range of temperature encompassing the physiological one (observed above 30 °C at low surface pressure). This phase behavior broadens the view of ceramide as a type of lipid not-always-rigid but able to exhibit polymorphic properties.     

Measurement of stray neutron doses inside the treatment room from a proton pencil beam scanning system

PurposeTo measure the environmental doses from stray neutrons in the vicinity of a solid slab phantom as a function of beam energy, field size and modulation width, using the proton pencil beam scanning (PBS) technique.MethodMeasurements were carried out using two extended range WENDI-II rem-counters and three tissue equivalent proportional counters. Detectors were suitably placed at different distances around the RW3 slab phantom. Beam irradiation parameters were varied to cover the clinical ranges of proton beam energies (100–220 MeV), field sizes ((2 × 2)–(20 × 20) cm²) and modulation widths (0–15 cm).ResultsFor pristine proton peak irradiations, large variations of neutron H¹(10)/D were observed with changes in beam energy and field size, while these were less dependent on modulation widths. H¹(10)/D for pristine proton pencil beams varied between 0.04 μSv Gy⁻¹ at beam energy 100 MeV and a (2 × 2) cm² field at 2.25 m distance and 90° angle with respect to the beam axis, and 72.3 μSv Gy⁻¹ at beam energy 200 MeV and a (20 × 20) cm² field at 1 m distance along the beam axis.ConclusionsThe obtained results will be useful in benchmarking Monte Carlo calculations of proton radiotherapy in PBS mode and in estimating the exposure to stray radiation of the patient. Such estimates may be facilitated by the obtained best-fitted simple analytical formulae relating the stray neutron doses at points of interest with beam irradiation parameters.     

Iron levels in the human brain: A post-mortem study of anatomical region differences and age-related changes

The link between brain iron homeostasis and neurodegenerative disease has been the subject of extensive research. There is increasing evidence of iron accumulation during ageing, and altered iron levels in some specific brain regions in neurodegenerative disease patients have been reported.Using graphite furnace atomic absorption spectrometry after microwave-assisted acid digestion of the samples, iron levels were determined in 14 different areas of the human brain [frontal cortex, superior and middle temporal, caudate nucleus, putamen, globus pallidus, cingulated gyrus, hippocampus, inferior parietal lobule, visual cortex of the occipital lobe, midbrain, pons (locus coeruleus), medulla and cerebellum (dentate nucleus)] of n = 42 adult individuals (71 ± 12 years old, range: 53–101 years old) with no known history or evidence of neurodegenerative, neurological or psychiatric disorders.It was found that the iron distribution in the adult human brain is quite heterogeneous. The highest levels were found in the putamen (mean ± SD, range: 855 ± 295 μg/g, 304–1628 μg/g) and globus pallidus (739 ± 390 μg/g, 225–1870 μg/g), and the lowest levels were observed in the pons (98 ± 43 μg/g, 11–253 μg/g) and medulla (56 ± 25 μg/g, 13–115 μg/g).Globally, iron levels proved to be age-related. The positive correlation between iron levels and age was most significant in the basal ganglia (caudate nucleus, putamen and globus pallidus).Compared with the age-matched control group, altered iron levels were observed in specific brain areas of one Parkinson's disease patient (the basal ganglia) and two Alzheimer's disease patients (the hippocampus).     

Biodegradability and mechanical properties of polycaprolactone composites encapsulating phosphate-solubilizing bacterium Bacillus sp. PG01

This study examined the feasibility of using polycaprolactone (PCL) and its composites (with starch and/or clay) in encapsulating cells of phosphate-solubilizing bacteria (PSB) for the development of biodegradable and “controlled-release” bacterial fertilizer. The PSB used in this work was an indigenous Bacillus sp. PG01 isolate. The results show that the PG01 strain was able to degrade all the cell-loaded capsules made of PCL and PCL composites, resulting in a continual cell release. Morphology observation indicates that severe disruption of the capsule structure occurred after incubation for 15–20 days. The biodegradability of the capsules decreased in the order of PCL/starch (20 wt%) > PCL/starch (20 wt%)/cay (7 wt%) > PCL alone > PCL/clay (7 wt%). Similar trends were also observed for the decrease in tensile strength and elongation at break, suggesting strong connections between biodegradability and the mechanical properties. Addition of starch appeared to enhance the biodegradability of the capsules, whereas the clay-blended composites were less biodegradable. The amount and rate of cell release from cell-encapsulated PCL-based capsules were positively dependent on the biodegradability and on the decrease in the mechanical strength. Nevertheless, the pattern of cell release was quite similar for all types of capsules. The outcome of this work seems to suggest that by proper manipulation of composite compositions, the controlled release of the bacterial fertilizer (i.e., Bacillus sp. PG01 cells) might be achievable.     

Low-temperature effect on enzyme activities involved in sucrose–starch partitioning in salt-stressed and salt-acclimated cotyledons of quinoa (Chenopodium quinoa Willd.) seedlings

The effect of low temperature on growth, sucrose–starch partitioning and related enzymes in salt-stressed and salt-acclimated cotyledons of quinoa (Chenopodium quinoa Willd.) was studied. The growth of cotyledons and growing axes in seedlings grown at 25/20 °C (light/dark) and shifted to 5/5 °C was lower than in those only growing at 25/20 °C (unstressed). However, there were no significant differences between low-temperature control and salt-treated seedlings. The higher activities of sucrose phosphate synthase (SPS, EC 2.4.1.14) and soluble acid invertase (acid INV, EC 3.2.1.25) were observed in salt-stressed cotyledons; however, the highest acid INV activity was observed in unstressed cotyledons. ADP-glucose pyrophosphorylase (ADP-GPPase, EC 2.7.7.27) was higher in unstressed cotyledons than in stressed ones. However, between 0 and 4 days the highest value was observed in salt-stressed cotyledons. The lowest value of ADP-GPPase was observed in salt-acclimated cotyledons. Low temperature also affected sucrose synthase (SuSy, EC 2.4.1.13) activity in salt-treated cotyledons. Sucrose and glucose were higher in salt-stressed cotyledons, but fructose was essentially higher in low-temperature control. Starch was higher in low-temperature control; however, the highest content was observed at 0 day in salt-acclimated cotyledons. Results demonstrated that low temperature induces different responses on sucrose–starch partitioning in salt-stressed and salt-acclimated cotyledons. Data also suggest that in salt-treated cotyledons source–sink relations (SSR) are changed in order to supply soluble sugars and proline for the osmotic adjustment. Relationships between starch formation and SuSy activity are also discussed.     

Osmoregulation in Dunaliella,Part II: Photosynthesis and starch contribute carbon for glycerol synthesis during a salt stress in Dunaliella tertiolecta

In response to an osmotic stress, Dunaliella tertiolecta osmoregulates by metabolizing intracellular glycerol as compatible solute. Upon the application of a salt stress to 0.17 M or 0.7 M NaCl grown D. tertiolecta cells, rates of total glycerol synthesis were substantially higher than that arising from photosynthetic ¹⁴CO₂ fixation into glycerol. The source of this extra carbon is the reserve starch pool. The contribution of carbon from the starch breakdown to glycerol synthesis was estimated from the difference between the total glycerol synthesized and that arising from ¹⁴CO₂ fixation. The maximum observed flux of carbon from ¹⁴CO₂ to glycerol from photosynthesis was of the order of 15–20 μmol ¹⁴C-glycerol mg⁻¹ Chl h⁻¹, whereas the total glycerol synthesis reached about 70 μmol glycerol mg⁻¹ Chl h⁻¹. The contribution of products of starch breakdown to glycerol synthesis increased progressively with increasing salt stress. In light, contrary to prevailing assumptions, both the photosynthesis and the starch breakdown contribute carbon to glycerol biosynthesis. The relative contributions of these two processes in the light, while cells were actively photosynthesizing, depended on the magnitude of the salt stress. On application of dilution stress, the flux of carbon from newly photosynthetically fixed ¹⁴CO₂ into glycerol was reduced progressively with increasing dilution stress that was also accompanied by a decline in total glycerol contents of the cell. The maximum observed rate of glycerol dissimilation was about 135 μmol glycerol mg⁻¹ Chl h⁻¹.     

Hydrolysis of starches by the action of an α-amylase from Bacillus subtilis

A moderately thermophilic Bacillus subtilis strain, isolated from fresh sheep’s milk, produced extracellular thermostable α-amylase. Maximum amylase production was obtained at 40 °C in a medium containing low starch concentrations. The enzyme displayed maximal activity at 135 °C and pH 6.5 and its thermostability was enhanced in the presence of either calcium or starch. This thermostable α-amylase was used for the hydrolysis of various starches. An ammonium sulphate crude enzyme preparation as well as the cell-free supernatant efficiently degraded the starches tested. The use of the clear supernatant as enzyme source is highly advantageous mainly because it decreases the cost of the hydrolysis. Upon increase of reaction temperature to 70 °C, all substrates exhibited higher hydrolysis rates. Potato starch hydrolysis resulted in a higher yield of reducing sugars in comparison to the other starches at all temperatures tested. Soluble and rice starch took, respectively, the second and third position regarding reducing sugars liberation, while the α-amylase studied showed slightly lower affinity for corn starch and oat starch.