Silks play a crucial role in the survival and reproduction of many insects. Labial glands, Malpighian tubules, and a variety of dermal glands have evolved to produce these silks. The glands synthesize silk proteins, which become semicrystalline when formed into fibers. Although each silk contains one dominant crystalline structure, the range of molecular structures that can form silk fibers is greater than any other structural protein group. On the basis of silk gland type, silk protein molecular structure, and the phylogenetic relationship of silk-producing species, we grouped insect silks into 23 distinct categories, each likely to represent an independent evolutionary event. Despite having diverse functions and fundamentally different protein structures, these silks typically have high levels of protein crystallinity and similar amino acid compositions. The substantial crystalline content confers extraordinary mechanical properties and stability to silk and appears to be required for production of fine protein fibers.

We isolated a bacterial strain, Agrobacterium radiobacter P230, which can hydrolyze a wide range of organophosphate (OP) insecticides. A gene encoding a protein involved in OP hydrolysis was cloned from A. radiobacter P230 and sequenced. This gene (called opdA) had sequence similarity to opd, a gene previously shown to encode an OP-hydrolyzing enzyme in Flavobacterium sp. strain ATCC 27551 and Brevundimonas diminuta MG. Insertional mutation of the opdA gene produced a strain lacking the ability to hydrolyze OPs, suggesting that this is the only gene encoding an OP-hydrolyzing enzyme in A. radiobacter P230. The OPH and OpdA proteins, encoded by opd and opdA, respectively, were overexpressed and purified as maltose-binding proteins, and the maltose-binding protein moiety was cleaved and removed. Neither protein was able to hydrolyze the aliphatic OP malathion. The kinetics of the two proteins for diethyl OPs were comparable. For dimethyl OPs, OpdA had a higher k cat than OPH. It was also capable of hydrolyzing the dimethyl OPs phosmet and fenthion, which were not hydrolyzed at detectable levels by OPH.Synthetic organophosphates (OPs) are used widely as insecticides in agriculture. OPs contain three phosphoester linkages and are hence often termed phosphotriesters. The phosphorus is also linked by a double bond to either an oxygen (PAO) in oxon OPs or a sulfur (PAS) in thion OPs. These insecticides are potent acetylcholinesterase (AchE) inhibitors, and various clinical effects can occur due to OP poisoning in humans. In general, hydrolysis of one of the phosphoester bonds reduces the toxicity of an OP, and in the case of parathion (O,O-diethyl p-nitrophenyl phosphorothioate), a 100-fold reduction in toxicity occurs (37,38).Enzymatic detoxification of OPs has become the focus of many studies because other means of removing OP residues are impractical or costly or are themselves environmentally hazardous. Enzymes from insect species resistant to OPs have been identified and considered for use in bioremediation (27). However, these enzymes are capable of hydrolyzing only oxon OPs and function at rates several orders of magnitude below the diffusion-limited maximum rate (28). Bacterial enzymes have also received considerable attention and may have advantages in terms of broader substrate specificities (both oxon and thion OPs) and superior kinetics (10, 13). The most widely studied bacterial enzyme is the OPH (organophosphorus-hydrolyzing) protein.OPH is a zinc-containing homodimeric protein found in the membrane of Flavobacterium sp. strain ATCC 27551 and Brevundimonas diminuta MG (5, 13). OPH is capable of hydrolyzing a wide range of oxon and thion OPs (13) and hydrolyzes paraoxon at a rate approaching the diffusion limits (6, 35). The OPH enzyme is encoded by the opd gene. Other opd-containing organisms (for example, a Pseudomonas strain) have been identified by using this gene in Southern hybridization analysis (8), while other OP-hydrolyzing organisms clearly do not contain the opd gene (10, 11). Flavobacterium sp...

An endosulfan-degrading mixed bacterial culture was enriched from soil with a history of endosulfan exposure. Enrichment was obtained by using the insecticide as the sole source of sulfur. Chemical hydrolysis was minimized by using strongly buffered culture medium (pH 6.6), and the detergent Tween 80 was included to emulsify the insecticide, thereby increasing the amount of endosulfan in contact with the bacteria. No growth occurred in control cultures in the absence of endosulfan. Degradation of the insecticide occurred concomitant with bacterial growth. The compound was both oxidized and hydrolyzed. The oxidation reaction favored the alpha isomer and produced endosulfate, a terminal pathway product. Hydrolysis involved a novel intermediate, tentatively identified as endosulfan monoaldehyde on the basis of gas chromatography-mass spectrometry and chemical derivatization results. The accumulation and decline of metabolites suggest that the parent compound was hydrolyzed to the putative monoaldehyde, thereby releasing the sulfite moiety required for growth. The monoaldehyde was then oxidized to endosulfan hydroxyether and further metabolized to (a) polar product(s). The cytochrome P450 inhibitor, piperonyl butoxide, did not prevent endosulfan oxidation or the formation of other metabolites. These results suggest that this mixed culture is worth investigating as a source of endosulfan-hydrolyzing enzymes for use in enzymatic bioremediation of endosulfan residues.

Vpu is a small phosphorylated integral membrane protein encoded by the human immunodeficiency virus type 1 genome and found in the endoplasmic reticulum and Golgi membranes of infected cells. It has been linked to roles in virus particle budding and degradation of CD4 in the endoplasmic reticulum. However, the molecular mechanisms employed by Vpu in performance of these functions are unknown. Structural similarities between Vpu and the M2 protein of influenza A virus have raised the question of whether the two proteins are functionally analogous: M2 has been demonstrated to form cation-selective ion channels in phospholipid membranes. In this paper we provide evidence that Vpu, purified after expression in Escherichia coli, also forms ion channels in planar lipid bilayers. The channels are approximately five-to sixfold more permeable to sodium and potassium cations than to chloride or phosphate anions. A bacterial cross-feeding assay was used to demonstrate that Vpu can also form sodium-permeable channels in vivo in the E. coli plasma membrane.

The effect of growth and carbohydrate production by the diatom Nitzschia cuwilineata on sediment erodibility was explored in laboratory flume experiments. Diatom cultures, incubated on sediment, were monitored daily for chlorophyll and carbohydrate concentrations and eroded in a recirculating flume at successive stages of growth. Because variations in erosion rate were far greater than variations in erosion threshold during the diatom growth period, erosion rate may be a more sensitive index of sediment stability. Erosion rate was negatively correlated with sediment chlorophyll (r2 = 0.759; P = 0.024) and bulk carbohydrate (r2 = 0.958; P = 0.001) concentrations. A strong negative correlation was found between the sediment bulk carbohydrate-to-chlorophyll ratio and erosion rate (r2 = 0.996; P < O.OOl), suggesting that this ratio would serve as a good indicator of sediment erodibility. The size of eroding particles or aggregates increased with the age of the biofilm, probably due to the pervasion of sediment with mucilage and changes in "stickiness" of the resulting sediment microfabric. Sediment stability increased throughout the stationary phase of growth regardless of lift forces produced by trapped bubbles within the biofilm and progressively smaller increases in sediment carbohydrate concentrations during this period. Knowledge of the physiological status of diatom biofilms is essential for the quantitative prediction of sediment transport, since diatom growth phase alters the behavior of sediment erosion.