Applications of Hydrolytic and Decarboxylating Enzymes in Biotransformations

Peptides, and oligosaccharides and glycosides, can be synthesised by making use of the ‘reverse hydrolytic activity’ of proteases and glycosidases respectively. In applying these enzymes to the practical synthesis of these classes of compound, several factors need to be considered, namely the need to shift the rate-determining step through the use of activated substrates, the need to minimise competing hydrolysis of these and the need to minimise hydrolysis of the products. In spite of these problems, the enzymatic methods have many attractive features, not least amongst which is the absolute control of stereochemistry in acyl transfer and glycosyl transfer respectively.     

Nature and properties of humic acid prepared from different sources and its effect on nutrient availability

Summary Effect of sources, moisture levels and extractants on extraction, analytical properties, IR-spectra, macromolecular characteristics, interaction with cations and physiological properties of the extracted material (humic acid) were studied. Among the three variables, extractant influenced the degree of humification, IR-spectra, nature and properties of humic acids to a greater extent though the other two had significant contribution. Sodium pyrophosphate found to be an ideal extractant for humic acid. Reduced viscosity of humic acids increased with dilution particularly below 0.1 g/dl and decreased with increasing H⁺ concentration between pH 6.5 and 8.5 irrespective of source, moisture level and extractant. The amount of proton released and drop in initial pH of humic acids on addition of different metal ions indicated formation of mono-, di-hydroxy metal-HA complexes which broke down at pH 8.0 and above. HA-Fe²⁺ and HA-Cu²⁺ complexes were found to be most stable. Humic acids from different sources varied significantly in their effect on available nutrient status of soils though, the latter is main determining factor particularly for calcium and magnesium.     

腐殖酸对小麦抗旱性的生理效应

在干旱气候条件下,喷施腐殖酸钠（ＨＡ）、黄腐酸钠（ＦＡ）可降低土壤水分损耗,提高小麦叶片持水能力,叶片细胞超氧物歧化酶（ＳＯＤ）、过氧化氢酶（ＣＡＴ）活性明显提高,丙二醛（ＭＤＡ）含量和电解质渗出率明显降低,可减缓叶绿素降解,增强光合速率和光合产物积累,延缓植株衰老,小麦抗旱性增强,千粒重增加．     

Habitat characteristics and the effect of various nutrient solutions on growth and mineral nutrition ofArnica montana L. grown on natural soil

Arnica montana, one of the character species of the replacement plant communityViolion caninae on sandy acid humic podzol, is declining in the Netherlands since 1950. Locally, it is even extinct.This process of decline may be attributed to (i) autonomic succession; (ii) increased rate of acidification of the soil and/or (iii) change in competitive relations amongArnica and more productive plant species, such as the successive dominantDeschampsia flexuosa. This paper examines the last two hypothesized factors, supposedly being regulated by atmospheric input of N-input, by (a) describing the habitat characteristics of a seemingly still healthy individual-rich population ofArnica and (b) growingArnica andDeschampsia in mixed humic podzol soil (mainly A₁ horizon) fertilized with variously-composed nutrient solutions in order to assess the nutrient supplying capacity of the podzol soil and species-specific nutritional demands related to their respective growth capabilities.The results suggest that an increased rate of soil acidification yielding extra supply of ionic Al and Mn is presumably of less importance. The implications of growth rate differences amongArnica andDeschampsia as related to their nutritional demands are likely far more important.Arnica grows more slowly thanDeschampsia in fertilized humus podzol. The latter species has a much more efficient use of nutrients. Furthermore, both species differ in K-nutrition if NH₄ ⁺ is the dominant N-source, a situation normally occurring in acid podzolic soils.Deschampsia possesses the capability to adapt its metabolic nutrition (avoiding nutritional imbalances) to a wide range of nutrient supplies thereby maintaining a rather constant level of growth.Arnica, on the contrary, lacks this capability. These results are discussed in the framework of competitive relations among co-occurring plant species in the plant communityViolion caninae. It is concluded that maintenance ofArnica and likely other character species of that alliance — all characterized by a low growth rate — will only be achieved when the plant community is properly managed by trampling, mowing or light levels of grazing (low stocking rate). Of prime importance is to maintain a low and relatively open vegetation structure at a relatively low level of nutrient supply.     

Contribution of bacterial cell nitrogen to soil humic fractions

Summary Living cells ofSerratia marcescens, uniformly labelled with¹⁵N, were added to samples of maple (Acer saccharum) and black spruce (Picea mariana) forest soils. After different periods of incubation from zero time to 100 days, the soils were subjected to alkali-acid and phenol extraction to provide humic acid, fulvic acid, humin and humoprotein fractions. Significant amounts of the cell nitrogen were recovered in the humic and fulvic acids immediately after addition. After incubation, less cell, nitrogen appeared in the humic acid and more in the fulvic acid. The amount of cell nitrogen recovered in the humin fraction increased with incubation. Roughly 5 to 10 per cent of the added cell nitrogen was found as amino acid nitrogen from humoprotein in a phenol extract of the humic acid. The data are consistent with the occurrence of co-precipitation of biologically labile biomass nitrogen compounds with humic polymers during the alkaline extraction procedure involved in the humic-fulvic fractionation.     

On the possible role of organic melanoidin polymers as matrices for prebiotic activity

Summary One of the major diagenetic pathways of organic matter in recent sediments involves the condensation of cellular constituents, particularly amino acids and sugars, into insoluble melanoidin-type polymers. These polymers consist mainly of humic and fulvic acids and make up the major part of the organic carbon reservoir in recent sediments. We suggest that a similar set of reactions between abiotically formed amino acids and sugars, and more generally between aldehydes and amines, occurred on a large scale in the prebiotic hydrosphere. The rapid formation of this insoluble polymeric material would have removed the bulk of the dissolved organic carbon from the primitive oceans and would thus have prevented the formation of an "organic soup".Melanoidin polymers have several properties which make them attractive hypothetical precursors of contemporary oxidation-reduction coenzymes: 1. they contain heterocyclic nitrogen compounds similar to the nitrogenous bases; 2. they contain a high concentration of stable free radicals; and 3. they tend to concentrate those heavy metals which play prominent roles in contemporary enzymic redox processes. The prebiotic formation of similar polymers could, therefore, have provided the starting point for a basic class of biochemical reactions.We suggest that the prebiotic scenario involved chemical and protoenzymic reactions at the sediment-ocean interface in relatively shallow waters and under conditions not much different from those of the recent environment.On leave from the Isotope Department, Weizmann Institute of Sciences, Rehovot, Israel. This is the address for reprint requests.On leave from the Department of Cell and Molecular Biology, San Francisco State University, San Francisco, California.On leave from the Department of Biophysics, University of Houston, Houston, Texas.     

The effect of pH on the lability of lead and cadmium sorbed on humic acid particles

Abstract

Sorption of lead(II) or cadmium(II) ions on humic acid particles (at pH 3.5) yields surface complexes which are sparingly soluble in the pH 3.55 region. Interaction of these species with acetic acid or dilute nitric acid released ASV labile metal species (mainly hydrated metal ion). When the pH was adjusted between 4 and 7, increasing amounts of the metal humate species (and humic acid substrate) dissolved and low levels of ASV labile species were detected. Overnight interaction with an excess of acetate ions (0.01 M) led to higher lability values (attributed to acetate/humate ligand exchange). The percentage of labile metal ion detected in acetate solutions varied with the amount of metal ion initially sorbed (range 100 to 500 mmol g^?1). With the lower loadings, the ASV labile levels peaked in the pH 6–7 region (at around 12% of total sorbed cadmium, and 4% of total lead). In alkaline solutions, the percentage of labile Cd fell to about half of the peak value, but with Pb, lability values increased at pH > 8, due possibly to the formation of hydroxy complex species. The ASV lability of the Cd and Pb humates, and their 24 hour lability values (determined using a transfer to cation exchanger technique) has been compared with the behaviour of Cu and Zn humates (using similar conditions).     

Metabolic profile and antioxidant responses during drought stress recovery in sugarcane treated with humic acids and endophytic diazotrophic bacteria

Water deficit is the major yield‐limiting factor for sugarcane crop production that can be enhanced by inoculating with plant growth promoting bacteria (PGPB) combined with humic substances. The aim of this work was to examine changes to the metabolic profile and antioxidant enzyme activity of sugarcane treated with PGPB and humic acid (HA) after drought and then rehydration. The drought was imposed by withholding irrigation for 21 days thereby measuring enzyme activity, metabolic profile and photosynthetic rate 1 week after rehydratation. Growth of plants treated with HA, PGPB and with both treatments combined (PGPB + HA) was higher than control plants. The antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) activities remained higher after rehydration only in plants treated with HA. Plants treated with HA and PGPB + HA exhibited increased transpiration, stomatal conductance and net photosynthesis than plants treated with PGPB. The PGPB‐treated plants exhibited drought resistance that resembled ‘delayed stress onset’, which is a new term for preserving water in the plants tissues. Water preservation in plants treated with PGPB was corroborated by higher relative water content (RWC) than control plants at the end of the drought period. Plants treated with HA + PGPB exhibited the highest water potential after rehydration and high RWC. Osmotic adjustment in the other treatments (control, HA and PGPB) was indicated by a new pattern of metabolic response after rehydration, including generally enhanced carbohydrates and proteins and specific changes induced by HA‐enhancing aromatic compounds, whereas PGPB exhibited enhanced fatty acids and other aliphatic H species. Humic acids assist with drought stress recovery by inducing antioxidant enzyme activity whereas PGPB induced preservation of leaf water potential and RWC by closing stomata efficiently, resulting in plant water preservation.     

Alleviation of salinity stress effects on agro-physiological traits of wheat by auxin,glycine betaine,and soil additives

Soil salinity is a major constraint to wheat production; it causes a severe reduction in wheat growth and yield. Alleviation of salinity effects on physiological, biochemical, and yield of wheat cultivars; Sids 14 and Misr 3 using some soil additions (control, Molasses and Humic acid), compatible solutes, and growth regulators (water as control, Naphthalene acetic acid, and Glycine betaine) were investigated in salt-affected soils. Results indicated that Misr 3 was superior to Sids 14 in all studied characteristics except flag leaf area, relative water content, plant height and recorded lower and desirable value of leaf temperature. The addition of Molasses (24 L ha^?1) or Humic acid (12 L ha^?1) significantly increased physiological and biochemical characteristics. At the same time, flag leaf temperature, proline, and malondialdehyde (MDA) content were decreased, yield and its attributes also increased except No. kernel spike-1. Foliar spray of Naphthalene acetic acid (NAA) at 30 mg L^?1. or glycine betaine (GB 100 mM) also positively affected the studied characteristics, where Glycine betaine recorded the highest Relative water content and Fv/Fm. In contrast, NAA recorded the most increased Catalase (CAT) activity, and the Number of spikes m^?2 and insignificant differences were observed between them in grain yield. It could be recommended the cultivation of Misr 3 with Molasses and GB under saline soils.     

Extracellular enzyme activities during humic acid degradation by the white rot fungi Phanerochaete chrysosporium and Trametes versicolor

Abstract The relationship between humic acid biodegradation and extracellular lignin peroxidase and Mn-dependent peroxidase activities of two white rot fungi, Phanerochaete chrysosporium and Tranetes versicolor , reported to be lignin degraders, was examined. In experimental conditions promoting culture aeration, particularly with T. versicolor no extracellular peroxidase activity could be detected unless humic acids were included in the culture medium. In the presence of humic acids, appreciable enzymatic activities were determined in the culture filtrate of the two fungi. However, T. versicolor was a more effective degrader than P. chrysosporium , and mineralization assays on synthetic humic acids with culture filtrates showed the important role played by Mn²⁺. The surfactant properties of humic acids are suggested to be responsible for the increase of enzymatic activities.